WO2024077391A1 - Composés thiazolo[5,4-d]pyrimidine, compositions les comprenant et leurs utilisations - Google Patents

Composés thiazolo[5,4-d]pyrimidine, compositions les comprenant et leurs utilisations Download PDF

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WO2024077391A1
WO2024077391A1 PCT/CA2023/051356 CA2023051356W WO2024077391A1 WO 2024077391 A1 WO2024077391 A1 WO 2024077391A1 CA 2023051356 W CA2023051356 W CA 2023051356W WO 2024077391 A1 WO2024077391 A1 WO 2024077391A1
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compound
alkyl
substituted
unsubstituted
cycloalkyl
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PCT/CA2023/051356
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Pierre Louis Beaulieu
Eric Beaulieu
Emeline BENOIT
Joanne TAN
Yannick Rose
Michael DORE
Mukund GHAVRE
Doris SCHUETZ
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Universite De Montreal
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D513/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
    • C07D513/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains two hetero rings
    • C07D513/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00

Definitions

  • This disclosure generally relates to thiazolo[5,4-d]pyrimidine compounds, pharmaceutical compositions comprising the same and their use in the treatment and prevention of diseases characterized by dysregulation of the RAS-ERK pathway (e.g. cancer, RASopathies).
  • diseases characterized by dysregulation of the RAS-ERK pathway e.g. cancer, RASopathies.
  • RAS-RAF-MEK-ERK (RAS: rat sarcoma; RAF: rapidly accelerated fibrosarcoma; MEK: mitogen-activated protein kinase; ERK: extracellular signal-regulated kinase) signaling pathway (hereafter referred to as the RAS-ERK pathway) plays a critical role in transmitting proliferation signals generated by growth factor receptors from the plasma membrane to the nucleus.
  • the pathway is dysregulated in a large proportion of cancers by activation of receptor tyrosine kinases (RTKs) (e.g.
  • ERBB1 , ERBB2, FLT3, RET, KIT activation or inactivation of RAS regulators (SOS1 and NF1) as well as constitutively activating mutations in RAS genes (/-/-, K- and NRAS', overall 30% of cancers) or in the BRAF gene (8% of cancers).
  • SOS1 and NF1 RAS regulators
  • constitutively activating mutations in RAS genes /-/-, K- and NRAS', overall 30% of cancers
  • BRAF gene 8% of cancers.
  • the prevalence of KRAS mutations is especially high in pancreatic (>90%), colorectal (50%), and lung (30%) cancers.
  • BRAF mutations are found with notably high frequencies in malignant melanoma (70 %), thyroid cancer (40 %) and colorectal cancer (10 %) (mutation frequencies based on COSMIC (Catalogue Of Somatic Mutations In Cancer; Wellcome Trust Sanger Institute) release v95, November 24 th 2021).
  • RAS proteins are small GTPases that convey extracellular growth signals to intracellular effectors to control vital processes like cell differentiation, proliferation and survival (Nat. Rev. Cancer2003, 3, 459).
  • Physiological activation of RAS occurs at the plasma membrane after stimulation of RTKs, which leads to GTP loading of the GTPase and thus its activation.
  • Activated RAS interacts and activates a battery of effector molecules, with the RAF kinases being the most critical RAS interactors in the context of cancer development (Nature Rev. Drug Discov. 2014, 13, 828).
  • Oncogenic mutations at Glycine 12, Glycine 13 or Glutamine 61 in RAS isoforms lead to aberrant and constitutive signaling in human cancer (Nat. Rev. Cancer 2003, 3, 459) (COSMIC release v95, November 24 th 2021).
  • RAF proteins Downstream of RAS, mammalian cells express three RAF paralogs (ARAF, BRAF and CRAF) that share a conserved C-terminal kinase domain (KD) (Nat. Rev. Mol. Cell Biol. 2015, 16, 281) and an N-terminal regulatory region (NTR) comprising a RAS-binding domain (RBD).
  • KD C-terminal kinase domain
  • NTR N-terminal regulatory region
  • RAF proteins are sequestered in the cytoplasm as monomers. Binding of GTP- bound activated RAS to the RBD induces membrane anchoring of RAF kinases (Nat. Rev. Mol. Cell Biol. 2015, 16, 281).
  • RAF proteins undergo kinase domain side-to-side dimerization and catalytic activation (Nature 2009, 461, 542).
  • Activated RAF proteins convey signals through a phosphorylation cascade from RAF to MEK and then MEK to ERK, leading to phosphorylation by ERK of an array of substrates eliciting cell-specific responses (Nat. Rev. Mol. Cell Biol. 2020, Oct., 21 (10), 607).
  • BRAF V600E valine to glutamic acid substitution at position 600
  • Cell 2004, 116, 855 the active form of BRAF
  • a diverse set of mutations occur at other residues (e.g. G466V, D594G, etc.) that lead to increased RAF signaling through a variety of mechanisms (Nat. Rev. Mol. Cell Biol. 2015, 16, 281).
  • BRAF V600E inhibitors are ineffective and even contraindicated against RAS-driven cancers.
  • the underlying mechanism results from the compounds’ ability to promote RAF kinase domain dimerization in the presence of active RAS (Nature 2010, 464, 431). This event is not restricted to BRAF, but also involves other RAF family members and is dictated by the compound binding mode and affinity (Nat. Chem. Biol. 2013, 9, 428).
  • the second strategy consisted in designing compounds that conformationally bias the BRAF kinase domain in the inactive state and thus do not paradoxically induce ERK signaling.
  • This has given rise to the “Paradox Breaker” (PB) molecule PLX8394, a derivative of PLX4032/vemurafenib (Nature 2015, 526, 583).
  • PB Paradox Breaker
  • PLX8394 a derivative of PLX4032/vemurafenib
  • the present technology relates to a compound of Formula I:
  • R 1 is selected from substituted or unsubstituted OR 3 , SR 3 , NH 2 , NHR 3 , N(R 3 ) 2 , C 3- 8 cycloalkyl, C 4-8 heterocycloalkyl, C 6-10 aryl and C 5-10 heteroaryl;
  • R 2 is selected from substituted C 6 aryl or C 5-10 heteroaryl, substituted or unsubstituted C 4- 8 heterocycloalkyl, and N(R 3 ) 2 ;
  • R 3 is independently in each occurrence selected from substituted or unsubstituted C 1-3 alkyl, C 3-8 cycloalkyl, C 4-8 heterocycloalkyl, C 6-10 aryl and C 5-10 heteroaryl;
  • X 1 is halo or an electron-withdrawing group
  • X 2 is selected from H, halo, and an electron-withdrawing group
  • X 3 and X 4 are each selected from H, halo, an electron-withdrawing group, C 1-3 alkyl, C 3- 4 cycloalkyl, and OC 1-3 alkyl; or a pharmaceutically acceptable salt or solvate thereof.
  • the present technology relates to a pharmaceutical composition for a use as defined in any one of the aforementioned embodiments, the composition comprising a compound as herein defined together with a pharmaceutically acceptable carrier, diluent or excipient.
  • the present technology relates to the use of a compound as herein defined for the treatment of a disease or disorder selected from a proliferative disease or disorder, a developmental anomaly caused by dysregulation of the RAS-ERK signaling cascade (RASopathies), or an inflammatory disease or an immune system disorder.
  • a disease or disorder selected from a proliferative disease or disorder, a developmental anomaly caused by dysregulation of the RAS-ERK signaling cascade (RASopathies), or an inflammatory disease or an immune system disorder.
  • the present technology also further relates to a method for the treatment of a disease or disorder selected from a proliferative disease or disorder, a developmental anomaly caused by dysregulation of the RAS-ERK signaling cascade (RASopathies), or an inflammatory disease or an immune system disorder, comprising administering a compound as herein defined to a subject in need thereof.
  • a method for inhibiting abnormal proliferation of cells comprising contacting the cells with a compound as defined herein is also contemplated.
  • the disease or disorder is selected from a neoplasm and a developmental anomaly, for instance, a disease or disorder associated with a RAF gene mutation (e.g. ARAF, BRAF or CRAF), a disease or disorder associated with a RAS gene mutation (e.g. KRAS), or a disease or disorder associated with both a RAF gene mutation and a RAS gene mutation.
  • a disease or disorder associated with a RAF gene mutation e.g. ARAF, BRAF or CRAF
  • a disease or disorder associated with a RAS gene mutation e.g. KRAS
  • the disease or disorder is associated with a receptor tyrosine kinase mutation or amplification (e.g. EGFR, HER2) or a mutation in a regulator of RAS downstream of the receptor (e.g. SOS1 gain of function, NF1 loss of function.
  • the disease or disorder is a neoplasm, such as those selected from melanoma, thyroid carcinoma (e.g. papillary thyroid carcinoma), colorectal, ovarian, breast cancer, endometrial cancer, liver cancer, sarcoma, stomach cancer, pancreatic carcinoma, Barret's adenocarcinoma, glioma (e.g. ependymoma), lung cancer (e.g. non-small cell lung cancer), head and neck cancer, acute lymphoblastic leukemia, acute myelogenous leukemia, non-Hodgkin's lymphoma, and hairy-cell leukemia.
  • thyroid carcinoma e.g. papillary thyroid carcinoma
  • colorectal ovarian
  • breast cancer endometrial cancer
  • liver cancer e.g. sarcoma
  • stomach cancer pancreatic carcinoma
  • Barret's adenocarcinoma e.g. ependymoma
  • lung cancer
  • the neoplasm is selected from colon or colorectal cancer, lung cancer, pancreatic cancer, thyroid cancer, breast cancer and melanoma.
  • any of the present uses and methods comprises inhibiting the RAS-ERK signaling pathway without substantial induction of a paradoxical pathway.
  • Figure 1 shows representative IC 50 inhibition dose response curves for compounds as described herein that do not induce paradoxical induction of pERK signaling (Y MIN >-20%) in RAS-mutant HCT116 cells (Examples 44 and 122) and a compound (PLX4720; CAS No. 918505-84-7) that causes strong induction of the pathway in the same cell line (Y MIN ⁇ -600%).
  • the term “about” or “approximately” means within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, i.e. , the limitations of the measurement system. For example, “about” can mean within 1 or more than 1 standard deviation, per the practice in the art. Alternatively, “about” can mean a range of up to 20%, preferably up to 10%, more preferably up to 5%, and more preferably still up to 1 % of a given value. Alternatively, particularly with respect to biological systems or processes, the term can mean within an order of magnitude, preferably within 5-fold, and more preferably within 2-fold, of a value. Where particular values are described in the application and claims, unless otherwise stated the term "about” meaning within an acceptable error range for the particular value should be assumed.
  • the terms “compounds”, “compounds herein described”, “compounds of the present application”, “thiazolo[5,4-d]pyrimidine compounds”, “thiazolopyrimidine compounds” and equivalent expressions refer to compounds described in the present application, e.g. those encompassed by structural Formula I, optionally with reference to any of the applicable embodiments, and also includes exemplary compounds, such as the compounds of Examples 1 to 159, as well as their pharmaceutically acceptable salts, solvates, esters, and prodrugs when applicable.
  • the compound may be drawn as its neutral form for practical purposes, but the compound is understood to also include its zwitterionic form.
  • Embodiments herein may also exclude one or more of the compounds.
  • Compounds may be identified either by their chemical structure or their chemical name. In a case where the chemical structure and chemical name would conflict, the chemical structure will prevail.
  • structures depicted herein are also meant to include all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational)) forms of the structure when applicable; for example, the R and S configurations for each asymmetric center. Therefore, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are within the scope of the present description.
  • the therapeutic compound unless otherwise noted, also encompasses all possible tautomeric forms of the illustrated compound, if any.
  • the term also includes isotopically labeled compounds where one or more atoms have an atomic mass different from the atomic mass most abundantly found in nature.
  • isotopes examples include, but are not limited to, 2 H (D), 3 H (T), 11 C, 13 C, 14 C, 15 N, 18 O, 17 O, any one of the isotopes of sulfur, etc.
  • the compound may also exist in unsolvated forms as well as solvated forms, including hydrated forms.
  • the compound may exist in multiple crystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated herein and are intended to be within the scope of the present invention.
  • a particular enantiomer may, in some embodiments be provided substantially free of the corresponding enantiomer and may also be enantiomerically enriched.
  • “Enantiomerically enriched” means that the compound is made up of a significantly greater proportion of one enantiomer. In certain embodiments the compound is made up of at least about 90% by weight of a preferred enantiomer. In other embodiments the compound is made up of at least about 95%, 98%, or 99% by weight of a preferred enantiomer.
  • Preferred enantiomers may be isolated from racemic mixtures by any method known to those skilled in the art, including high- pressure liquid chromatography (HPLC) on chiral support and the formation and crystallization of chiral salts or be prepared by asymmetric syntheses.
  • HPLC high- pressure liquid chromatography
  • pharmaceutically acceptable salt refers to those salts of the compounds of the present description which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio.
  • Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge, et al. describes pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19 (1977).
  • the salts can be prepared in situ during the final isolation and purification of the compounds of the present description, or separately by reacting a free base function of the compound with a suitable organic or inorganic acid (acid addition salts) or by reacting an acidic function of the compound with a suitable organic or inorganic base (base-addition salts).
  • solvate refers to a physical association of one of the present compound with one or more solvent molecules, including water and non-aqueous solvent molecules. This physical association may include hydrogen bonding. In certain instances, the solvate will be capable of isolation, for example when one or more solvent molecules are incorporated in the crystal lattice of a crystalline solid.
  • solvate encompasses both solution-phase and isolable solvates.
  • Exemplary solvates include, without limitation, hydrates, hemihydrates, ethanolates, hemiethanolates, n-propanolates, iso-propanolates, 1 -butanolates, 2-butanolate, and solvates of other physiologically acceptable solvents, such as the Class 3 solvents described in the International Conference on Harmonization (I CH), Guide for Industry, Q3C Impurities: Residual Solvents (1997). Accordingly, the compound as herein described also includes each of its solvates and mixtures thereof.
  • ester refers to esters of the compounds formed by the process of the present description which may hydrolyze in vivo and include those that break down readily in the human body to leave the parent compound or a salt thereof.
  • Suitable ester groups include, for example, those derived from pharmaceutically acceptable aliphatic carboxylic acids, particularly alkanoic, alkenoic, cycloalkanoic and alkanedioic acids, in which each alkyl or alkenyl moiety advantageously has not more than 6 carbon atoms.
  • esters include, but are not limited to, formates, acetates, propionates, butyrates, acrylates and ethylsuccinates of hydroxyl groups, and alkyl esters of an acidic group.
  • Other ester groups include sulfonate or sulfate esters.
  • prodrugs refers to those prodrugs of the compounds formed by the process of the present description which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals with undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and effective fortheir intended use.
  • Prodrug as used herein means a compound which is convertible in vivo by metabolic means (e.g. by hydrolysis) to afford any compound delineated by the formulae of the instant description.
  • the number of carbon atoms in a hydrocarbyl substituent can be indicated by the prefix “C x -C y " or "C x - y " where x is the minimum and y is the maximum number of carbon atoms in the substituent.
  • x and y are associated with a group incorporating one or more heteroatom(s) by definition (e.g. heterocycloalkyl, heteroaryl, etc).
  • x and y define respectively the minimum and maximum number of atoms in the cycle, including carbon atoms as well as heteroatom(s).
  • alkyl refers to a saturated, straight- or branched-chain hydrocarbon radical typically containing from 1 to 20 carbon atoms.
  • C 1-8 alkyl contains from one to eight carbon atoms.
  • alkyl radicals include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, neopentyl, n-hexyl, heptyl, octyl radicals and the like.
  • alkenyl denotes a straight- or branched-chain hydrocarbon radical containing one or more double bonds and typically from 2 to 20 carbon atoms.
  • C 2- 8 alkenyl contains from two to eight carbon atoms.
  • Alkenyl groups include, but are not limited to, for example, ethenyl, propenyl, butenyl, l-methyl-2-buten-l-yl, heptenyl, octenyl and the like.
  • alkynyl denotes a straight- or branched-chain hydrocarbon radical containing one or more triple bonds and typically from 2 to 20 carbon atoms.
  • C 2- 8 alkynyl contains from two to eight carbon atoms.
  • Representative alkynyl groups include, but are not limited to, for example, ethynyl,1-propynyl, 1-butynyl, heptynyl, octynyl and the like.
  • cycloalkyl refers to a group comprising a saturated or partially unsaturated (non aromatic) carbocyclic ring in a monocyclic or polycyclic ring system, including spiro (sharing one atom), fused (sharing at least one bond) or bridged (sharing two or more bonds) carbocyclic ring systems, having from three to fifteen ring members.
  • cycloalkyl groups include, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclopenten-1-yl, cyclopenten-2-yl, cyclopenten-3-yl, cyclohexyl, cyclohexen-1-yl, cyclohexen-2-yl, cyclohexen-3-yl, cycloheptyl, bicyclo[4,3,0]nonanyl, norbornyl, and the like.
  • the term cycloalkyl includes both unsubstituted cycloalkyl groups and substituted cycloalkyl groups.
  • C 0 - n cycloalkyl refers to a cycloalkyl group having from 3 to the indicated “n” number of carbon atoms in the ring structure. Unless the number of carbons is otherwise specified, “lower cycloalkyl” groups as herein used, have at least 3 and equal or less than 8 carbon atoms in their ring structure.
  • heterocycle As used herein, the terms “heterocycle”, “heterocycloalkyl”, “heterocyclyl”, “heterocyclic radical”, and “heterocyclic ring” are used interchangeably and refer to a chemically stable 3- to 7- membered monocyclic or 7-10-membered bicyclic heterocyclic moiety that is either saturated or partially unsaturated, and having, in addition to carbon atoms, one or more, preferably one to four, heteroatoms, as defined above.
  • nitrogen includes a substituted nitrogen.
  • the nitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or NR (as in N-substituted pyrrolidinyl).
  • a heterocyclic ring can be attached to its pendant group at any heteroatom or carbon atom that results in a chemically stable structure and any of the ring atoms can be optionally substituted.
  • heterocycloalkyl groups include, but are not limited to, 1 ,3-dioxolanyl, pyrrolidinyl, pyrrolidonyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiopyranyl, tetrahydrodithienyl, tetrahydrothienyl, thiomorpholino, thioxanyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepiny
  • Heterocyclic groups also include groups in which a heterocyclic ring is fused to one or more aryl, heteroaryl, or cycloaliphatic rings, such as indolinyl, 3H-indolyl, chromanyl, chromenyl, phenanthridinyl, 2- azabicyclo[2.2.1]heptanyl, octahydroindolyl, or tetrahydroquinolinyl, where the radical or point of attachment is on the heterocyclyl ring.
  • a heterocyclyl group may be mono- or bicyclic.
  • heterocyclylalkyl refers to an alkyl group substituted by a heterocyclyl, wherein the alkyl and heterocyclyl portions independently are optionally substituted.
  • C 3 - n heterocycloalkyl refers to a heterocycloalkyl group having from 3 to the indicated “n” number of atoms in the ring structure, including carbon atoms and heteroatoms.
  • partially unsaturated refers to a ring moiety that includes at least one double or triple bond between ring atoms but is not aromatic.
  • partially unsaturated is intended to encompass rings having multiple sites of unsaturation but is not intended to include aryl or heteroaryl moieties, as herein defined.
  • aryl used alone or as part of a larger moiety as in “aralkyl”, “aralkoxy”, “aryloxy”, or “aryloxyalkyl”, refers to aromatic groups having 4n+2 conjugated ⁇ (pi) electrons, wherein n is an integer from 1 to 3, in a monocyclic moiety or a bicyclic or tricyclic fused ring system having a total of six to 15 ring members, wherein at least one ring in the system is aromatic and wherein each ring in the system contains three to seven ring members.
  • aryl may be used interchangeably with the term “aryl ring”.
  • aryl refers to an aromatic ring system which includes, but not limited to, phenyl, biphenyl, naphthyl, azulenyl, anthracyl and the like, which may bear one or more substituents.
  • aralkyl or “arylalkyl” refers to an alkyl residue attached to an aryl ring. Examples of aralkyl include, but are not limited to, benzyl, phenethyl, and the like.
  • aryl is a group in which an aromatic ring is fused to one or more non-aromatic rings, such as indanyl, indenyl, phthalimidyl, naphthimidyl, fluorenyl, phenanthridinyl, or tetrahydronaphthyl, and the like.
  • C 6 - n aryl refers to an aryl group having from 6 to the indicated “n” number of atoms in the ring structure.
  • heteroaryl used alone or as part of a larger moiety, e.g., "heteroaralkyl”, or “heteroaralkoxy”, refers to aromatic groups having 4n+2 conjugated ⁇ (pi) electrons, wherein n is an integer from 1 to 3 (e.g. having 5 to 18 ring atoms, preferably 5, 6, or 9 ring atoms; having 6, 10, or 14 ⁇ electrons shared in a cyclic array); and having, in addition to carbon atoms, from one to five heteroatoms.
  • heteroatom includes but is not limited to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quaternized form of a basic nitrogen.
  • heteroaryl may be a single ring, or two or more fused rings.
  • heteroaryl also includes groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclic rings, where the radical or point of attachment is on the heteroaromatic ring.
  • heteroaryl groups include thienyl, furanyl (furyl), pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, indolyl, 3H-indolyl, isoindolyl, indolizinyl, benzothienyl (benzothiophenyl), benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzoxazolyl, benzothiazolyl, benzotriazolyl, pyrrolopyridinyl (e.g.
  • pyrrolo[3,2- b]pyridinyl or pyrrolo[3,2-c]pyridinyl) pyrazolopyridinyl (e.g. pyrazolo[1 ,5-a]pyridinyl), furopyridinyl, purinyl, imidazopyrazinyl (e.g.
  • imidazo[4,5-b]pyrazinyl quinolyl (quinolinyl), isoquinolyl (isoquinolinyl), quinolonyl, isoquinolonyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, naphthyridinyl, and pteridinyl carbazolyl, acridinyl, phenanthridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-l,4-oxazin-3(4H)-one.
  • a heteroaryl group may be mono- or bicyclic. Heteroaryl groups include rings that are optionally substituted.
  • the term "heteroaralkyl” refers to an alkyl group substituted by a heteroaryl, wherein the alkyl and heteroaryl portions independently are optionally substituted. Examples include, but are not limited to, pyridinylmethyl, pyrimidinylethyl and the like.
  • the term “C 5 - n heteroaryl” refers to a heteroaryl group having from 5 to the indicated “n” number of atoms in the ring structure, including carbon atoms and heteroatoms.
  • compounds of the present description may contain “optionally substituted” moieties.
  • substituted whether preceded by the term “optionally” or not, means that one or more hydrogens of the designated moiety are replaced with a suitable substituent.
  • an “optionally substituted” group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at each position.
  • Combinations of substituents envisioned under the present description are preferably those that result in the formation of chemically stable or chemically feasible compounds.
  • chemically stable refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, and, in certain embodiments, their recovery, purification, and use for one or more of the purposes disclosed herein.
  • halo designates a halogen atom, i.e. a fluorine, chlorine, bromine or iodine atom, preferably fluorine or chlorine.
  • optionally substituted refers to groups that are substituted or unsubstituted by independent replacement of one, two, or three or more of the hydrogen atoms thereon with substituents including, but not limited to F, Cl, Br, I, OH, CO 2 H, alkoxy, oxo, thiooxo, NO 2 , CN, CF 3 , NH 2 , NHalkyl, NHalkenyl, NHalkynyl, NHcycloalkyl, NHaryl, NHheteroaryl, NHheterocyclic, dialkylamino, diarylamino, diheteroarylamino, O-alkyl, O-alkenyl, O-alkynyl, O-cycloalkyl, O-aryl, O-heteroaryl, O-haloalkyl, O-heterocyclic, C(O)alkyl, C(O)alkenyl, C(O)alkynyl, C(
  • variable herein includes definitions of that variable as any single group or combination of listed groups.
  • the recitation of an embodiment for a variable herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof.
  • the recitation of an embodiment herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof. As such, the following embodiments are present alone or in combination if applicable.
  • the present compounds present a thiazolo[5,4-d]pyrimidine core structure to which is attached defined substituents to achieve the product’s beneficial activity.
  • Examples of thiazolopyrimidine compounds as defined herein are illustrated by general Formula I:
  • R 1 is selected from substituted or unsubstituted OR 3 , SR 3 , NH 2 , NHR 3 , N(R 3 ) 2 , C 3 - 8cycloalkyl, C 4-8 heterocycloalkyl, C 6-10 aryl and C 5-10 heteroaryl, for example, selected from substituted or unsubstituted SR 3 , N(R 3 ) 2 , C 4-8 heterocycloalkyl, C 6-10 aryl and C 5-10 heteroaryl, preferably a substituted or unsubstituted C 6-10 aryl or C 5-10 heteroaryl;
  • R 2 is selected from substituted Cearyl or C 5-10 heteroaryl, substituted or unsubstituted C 4 - 8heterocycloalkyl, and N(R 3 ) 2 ;
  • R 3 is independently in each occurrence selected from substituted or unsubstituted C 1-3 alkyl, C 3-8 cycloalkyl, C 4-8 heterocycloalkyl, C 6-10 aryl and C 5-10 heteroaryl;
  • X 1 is halo or an electron-withdrawing group
  • X 2 is selected from H, halo, and an electron-withdrawing group
  • X 3 and X 4 are each selected from H, halo, an electron-withdrawing group, C 1-3 alkyl, C 3 - 4cycloalkyl, and OCiwalkyl; or a pharmaceutically acceptable salt or solvate thereof.
  • the electron-withdrawing group is selected from perhaloalkyl (e.g. CF 3 or CCI 3 ), CN, NO 2 , sulfonate, alkylsulfonyl (e.g. SO 2 Me or SO 2 CF 3 ), alkylcarbonyl (e.g. C(O)Me), carboxylate, alkoxycarbonyl (e.g. C(O)OMe), and aminocarbonyl (e.g. C(O)NH 2 ).
  • X 1 is Cl and X 2 is F, or X 1 is F and X 2 is H, or X 1 and X 2 are both F.
  • X 3 and X 4 are each H.
  • X 3 is F and X 4 is H.
  • aminoarylsulfonamide moiety in Formula I may be designated L and preferably be selected from:
  • R 2 is a substituted Cearyl or C 5-10 heteroaryl, e.g. R 2 is a Cearyl substituted with at least one group selected from F, Cl, Br, CN, NO 2 , and a substituted or unsubstituted C 1- 8 alkyl, C 3-4 cycloalkyl or OC 1-3 alkyl.
  • R 2 is a group of the formula: R 4 wherein:
  • R 4 is selected from H, F, Cl, Br, CN, and a substituted or unsubstituted C 1-3 alkyl, C 3 - 4cycloalkyl or OC 1-3 alkyl, e.g. R 4 is selected from H, F, Cl, Br, Me, Et, CN, CHF2, and CF 3 ;
  • R 5 is selected from H, F, Cl, CN, and a substituted or unsubstituted C 1-3 alkyl, C 3-4 cycloalkyl, or OC 1-3 alkyl, e.g. R 5 is selected from H, F, Me, CF 3 , CN, and Cl;
  • R 6 is selected from H, F, Cl, Br, NO 2 , NH 2 , and a substituted or unsubstituted C 1-3 alkyl, C 3 - 4cycloalkyl or OC 1-3 alkyl, e.g. R 6 is selected from H, F, Cl, Br, and a substituted or unsubstituted C 1-3 alkyl, C 3-4 cycloalkyl or OC 1-3 alkyl, or R 6 is selected from H, F, Cl, Me, Et, and OMe;
  • R 7 is selected from H, F, Cl, and a substituted or unsubstituted C 1-3 alkyl, e.g. R 7 is selected from H, Me, F, and Cl;
  • R 8 is selected from H, F, and a substituted or unsubstituted C 1-3 alkyl, e.g. R 8 is selected from H, Me and F; or R 4 and R 5 or R 5 and R 6 are taken together with their adjacent carbon atoms to form a substituted or unsubstituted carbocycle or heterocycle provided that the heterocycle (R 2 ) is not a benzoxazolinone; and
  • ( — ) represents a bond serving as a point of attachment between R 2 and the rest of the molecule; wherein when R 4 is H or F, then at least one of R 5 , R 6 , R 7 or R 8 is other than H or F; and wherein when R 5 is CN, then at least one of R 4 , R 6 , R 7 or R 8 is other than H.
  • R 8 is H.
  • R 4 is selected from F, Cl, Et and Me
  • R 5 , R 7 , and R 8 are each H
  • R 6 is selected from H, Cl, Me and OMe.
  • R 4 is selected from F, Cl, and Me
  • R 6 , R 7 and R 8 are each H
  • R 5 is selected from F and Cl.
  • R 4 is selected from Cl and a substituted or unsubstituted C 1-3 alkyl (e.g. Me);
  • R 5 is selected from H, F, Cl, and a substituted or unsubstituted C 1-3 alkyl (e.g., Me);
  • R 6 is selected from H and a substituted or unsubstituted OC 1-3 alkyl (e.g. OCH 3 ); and
  • R 7 and R 8 are each H.
  • R 4 is selected from H, Cl, Br and methyl;
  • R 5 is selected from H, F, and Cl;
  • R 6 is selected from H, F, Cl, Me and OMe; and
  • R 7 and R 8 are each H.
  • R 4 is selected from Cl and a substituted or unsubstituted C 1-3 alkyl, preferably R 4 is Cl or Me;
  • R 5 is selected from H, F, Cl, and a substituted or unsubstituted C 1-3 alkyl (e.g., Me), preferably R 5 is F, Cl or Me;
  • R 6 is selected from H, F, Cl, a substituted or unsubstituted C 1-3 alkyl (e.g., Me), and a substituted or unsubstituted OC 1-3 alkyl (e.g.
  • R 6 is H or F, or R 6 is Cl or a substituted or unsubstituted C 1-3 alkyl or substituted or unsubstituted OC 1- 8 alkyl, or R 6 is CH 3 or OCH 3 ; and R 7 and R 8 are each H.
  • R 6 is a substituted C 1-3 alkyl.
  • R 2 is a substituted C 5 heteroaryl, such as a group of the formula: wherein:
  • X 5 is selected from NH, NC 1-3 alkyl, NC 3-4 cycloalkyl, O and S;
  • R 9 , R 10 , R 11 are each independently selected from H, F, Cl, CN, and a substituted or unsubstituted C 1-3 alkyl, C 3-4 cycloalkyl, C(O)OC 1-3 alkyl or OC 1-3 alkyl, provided that one of R 9 and R 11 is H and the other is not H; and
  • ( — ) represents a bond serving as a point of attachment between R 2 and the rest of the molecule.
  • R 2 is a group of the formula: wherein:
  • X 5 is selected from NH, NC 1-3 alkyl, NC 3-4 cycloalkyl, O and S;
  • R 9 is selected from F, Cl, CN, and a substituted or unsubstituted C 1-3 alkyl, C 3-4 cycloalkyl, C(O)OC 1-3 alkyl or OC 1-3 alkyl;
  • R 10 and R 12 are each independently selected from H, F, Cl, CN, and a substituted or unsubstituted C 1-3 alkyl, C 3-4 cycloalkyl, C(O)OC 1-3 alkyl or OC 1-3 alkyl; and
  • ( — ) represents a bond serving as a point of attachment between R 2 and the rest of the molecule.
  • R 9 and R 10 are each independently selected from F, Cl, CN, and a substituted or unsubstituted C 1-3 alkyl, C 3-4 cycloalkyl, C(O)OC 1-3 alkyl or OC 1-3 alkyl, preferably Cl and a substituted or unsubstituted C 1-3 alkyl, more preferably R 9 and R 10 are both Cl.
  • X 5 is O or S, preferably S.
  • R 2 is a substituted C 5-10 heteroaryl, such as a group of the formula: wherein:
  • X 9 , X 10 , X 11 , X 12 , and X 13 are independently selected from N and C, wherein at least one and at most two of X 9 , X 10 , X 11 , X 12 , and X 13 are N; and
  • R 19 , R 20 , R 21 , R 22 and R 23 are selected from H, F, Cl, Br, CN, NO 2 , NH 2 , and a substituted or unsubstituted C 1-3 alkyl, C 3-4 cycloalkyl or OC 1-3 alkyl, or are absent when their attached X 9 , X 10 , X 11 , X 12 , or X 13 is N; wherein at least one of X 9 and X 13 is not N; wherein when one of X 9 and X 13 is N, then the other is not N or CH; and
  • ( — ) represents a bond serving as a point of attachment between R 2 and the rest of the molecule.
  • R 2 is a C 5 heterocycloalkyl.
  • R 2 is a group of the formula: wherein:
  • R 13 is independently in each occurrence selected from F, Cl, and a substituted or unsubstituted C 1-3 alkyl, C 3-4 cycloalkyl, or C 1-3 alkoxy; n is an integer selected from 0 to 8; or n is between 2 and 8 and two R 13 are taken together with their adjacent carbon atoms to form a C 3-4 cycloalkyl; and
  • ( — ) represents a bond serving as a point of attachment between R 2 and the rest of the molecule.
  • R 13 is in the 3-position. In another embodiment, R 13 is selected from F, Me, OMe, and CH 2 OMe, and n is 1 or 2. For instance, R 13 is a methoxy group in the 3-position and n is 1.
  • R 2 is N(R 3 ) 2 .
  • R 2 is N(R 3 ) 2 and R 3 is selected from substituted or unsubstituted C 1-3 alkyl or C 3-8 cycloalkyl.
  • the compound of Formula I is a compound of Formula II, or a pharmaceutically acceptable salt or solvate thereof: wherein R 1 , R 4 , R 5 , and R 6 are each independently as defined herein, preferably R 4 is selected from Cl, Br and methyl; R 5 is selected from H, F, Cl and methyl; R 6 is selected from H, F, Cl, Me and OMe.
  • the compound of Formula I is a compound of Formula III, or a pharmaceutically acceptable salt or solvate thereof:
  • R 1 , R 9 , R 10 , R 12 , and X 5 are each independently as defined herein.
  • R 2 groups are illustrated by groups B1 to B77 defined as follows:
  • ( — ) represents a bond serving as a point of attachment between R 2 and the rest of the molecule.
  • R 2 is selected from groups B1 to B77, or preferably R 2 is selected from groups
  • R 1 is OR 3 or SR 3 , for instance, R 1 is SR 3 .
  • R 3 is a substituted or unsubstituted C 1-3 alkyl (e.g. C 1-3 alkyl).
  • R 1 is a substituted or unsubstituted C 6 aryl group. In another embodiment, R 1 is a substituted or unsubstituted C 4-6 heterocycloalkyl group. For instance, R 1 is a C 4- 5 heterocycloalkyl optionally substituted with one or two groups selected from halo, OH, C 1-6 alkyl, and OC 1-6 alkyl. For instance, R 1 is a /V-pyrrolidinyl group substituted with one or two groups selected from F and OH.
  • R 1 is a substituted or unsubstituted C 5-6 heteroaryl group or a substituted or unsubstituted Cgheteroaryl group.
  • R 1 is a substituted or unsubstituted group selected from thienyl, imidazolyl, pyrazolyl, triazolyl, thiazolyl, pyridyl, pyrimidinyl, indolyl, indazolyl, benzimidazolyl, benzotriazolyl, pyrrolopyridinyl (e.g.
  • pyrrolo[3,2-b]pyridinyl or pyrrolo[3,2- c]pyridinyl pyrazolopyridinyl (e.g. pyrazolo[1 ,5-a]pyridinyl), purinyl, and imidazopyrazinyl (e.g. imidazo[4,5-b]pyrazinyl), more preferably attached to the thiazolopyrimidine core through a nitrogen atom.
  • R 1 examples include a substituted or unsubstituted group selected from: wherein (— ) represents a bond serving as a point of attachment between R 1 and the rest of the molecule.
  • R 1 is a substituted or unsubstituted group selected from: wherein (— ) represents a bond serving as a point of attachment between R 1 and the rest of the molecule.
  • R 1 is one of the above groups further substituted with at least one substituent selected from OH, halo, CN, NO 2 , C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, OC 1-6 alkyl, C 5-10 heteroaryl, C 3-10 cycloalkyl, C 4-10 heterocycloalkyl, C(O)R 15 , C(O)N(R 14 ) 2 , SO 2 R 15 , SO 2 N(R 14 ) 2 , N(R 16 )C(O)R 15 , N(R 16 )SO 2 R 15 , N(R 16 )C(O)N(R 14 ) 2 , N(R 16 )SO 2 N(R 14 ) 2 , N(R 14 ) 2 , P(O)(R 15 ) 2 , CH 2 C(O)R 15 , CH 2 C(O)N(R 14 ) 2 , CH 2 SO 2 R 15 , CH 2 SO 2 N
  • R 14 is independently in each occurrence selected from H, C 1-6 alkyl, C 2-6 alkenyl, C 2- 6 alkynyl, C 3-10 cycloalkyl, C 4-10 heterocycloalkyl, Cearyl, and C 5-10 heteroaryl, or two R 14 are taken together with their adjacent nitrogen atom to form a C 4-10 heterocycloalkyl group;
  • R 15 is independently in each occurrence selected from C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-10 cycloalkyl, Cearyl, and C 5-10 heteroaryl;
  • R 16 is independently in each occurrence selected from H, C 1-6 alkyl, C 2-6 alkenyl, C 2- 6 alkynyl, C 3-10 cycloalkyl, Cearyl, and C 5-10 heteroaryl; wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group, included in R 1 (including in the definitions of R 14 , R 15 , and R 16 ), is optionally further substituted.
  • R 1 is a group of the formula: wherein:
  • R 17 is selected from H, OH, halo, CN, NO 2 , C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, OC 1-6 alkyl, C 5-10 heteroaryl, C 3-10 cycloalkyl, C 4-10 heterocycloalkyl, C(O)R 15 , C(O)N(R 14 ) 2 , SO 2 R 15 , SO 2 N(R 14 ) 2 , N(R 16 )C(O)R 15 , N(R 16 )SO 2 R 15 , N(R 16 )C(O)N(R 14 ) 2 , N(R 16 )SO 2 N(R 14 ) 2 , N(R 14 ) 2 , P(O)(R 15 ) 2 , CH 2 C(O)R 15 , CH 2 C(O)N(R 14 ) 2 , CH 2 SO 2 R 15 , CH 2 SO 2 N(R 14 ) 2 ,
  • R 27 is selected from H, OH, halo, CN, NO 2 , C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, OC 1-6 alkyl, C 5-10 heteroaryl, C 3-10 cycloalkyl, C 4-10 heterocycloalkyl, C(O)R 15 , C(O)N(R 14 ) 2 , SO 2 R 15 , SO 2 N(R 14 ) 2 , N(R 16 )C(O)R 15 , N(R 16 )SO 2 R 15 , N(R 16 )C(O)N(R 14 ) 2 , N(R 16 )SO 2 N(R 14 ) 2 , N(R 14 ) 2 , P(O)(R 15 ) 2 , CH 2 C(O)R 15 , CH 2 C(O)N(R 14 ) 2 , CH 2 SO 2 R 15 , CH 2 SO 2 N(R 14 ) 2 ,
  • CH 2 N(R 16 )C(O)R 15 CH 2 N(R 16 )SO 2 R 15 , CH 2 N(R 16 )C(O)N(R 14 ) 2 , CH 2 N(R 16 )SO 2 N(R 14 ) 2 , and CH 2 N(R 14 ) 2 , preferably H, halo (e.g. F), optionally substituted C 1-6 alkyl, or optionally substituted OC 1-6 alkyl;
  • X 6 is N or CH
  • X 7 is N and R 18 is absent;
  • X 7 is C and R 18 is selected from C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, OC 1-6 alkyl, C 5- wheteroaryl, C 3-10 cycloalkyl, C 4-10 heterocycloalkyl, C(O)R 15 , C(O)N(R 14 ) 2 , SO 2 R 15 , SO 2 N(R 14 ) 2 , N(R 16 )C(O)R 15 , N(R 16 )SO 2 R 15 , N(R 16 )C(O)N(R 14 ) 2 , N(R 16 )SO 2 N(R 14 ) 2 , N(R 14 ) 2 , P(O)(R 15 ) 2 , CH 2 C(O)R 15 , CH 2 C(O)N(R 14 ) 2 , CH 2 SO 2 R 15 , CH 2 SO 2 N(R 14 ) 2 ,
  • R 14 , R 15 , and R 16 are as defined above; wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, or heteroaryl, included in R 1 (including in the definitions of R 14 , R 15 , R 16 , R 17 and R 18 ), is optionally further substituted; and wherein (— ) represents a bond serving as a point of attachment between R 1 and the rest of the molecule.
  • R 1 is a group of the formula: wherein:
  • X 15 , X 16 , X 17 , and X 18 are independently selected from O, N, S, and CR 17 , wherein R 17 is as previously defined; wherein at most two of X 15 , X 16 , X 17 , and X 18 are O, N, or S; and wherein (— ) represents a bond serving as a point of attachment between R 1 and the rest of the molecule.
  • the compound of Formula I is a compound of Formula IV or V, or a pharmaceutically acceptable salt or solvate thereof:
  • R 4 , R 5 , R 6 , R 17 , R 18 , R 27 , X 6 , X 7 , X 15 , X 16 , X 17 , and X 18 are each independently as defined herein, preferably R 4 is selected from Cl, Br and methyl; R 5 is selected from H, F, Cl and methyl; R 6 is selected from H, Cl, F, Me and OMe.
  • the compound of Formula I is a compound of Formula VI or VII, or a pharmaceutically acceptable salt or solvate thereof: wherein R 9 , R 10 , R 12 , R 17 , R 18 , R 27 , X 5 , X 6 , X 7 , X 15 , X 16 , X 17 and X 18 are each independently as defined herein.
  • X 6 is N. In another embodiment, X 6 is CH. In another embodiment, X 7 is N, R 17 is selected from H, OH, halo, CN, C 1-6 alkyl, C 2-6 alkenyl, C 2- 6 alkynyl, OC 1-6 alkyl, C 5-10 heteroaryl, C 3-10 cycloalkyl, C 4-10 heterocycloalkyl, C(O)R 15 , C(O)N(R 14 ) 2 , SO 2 R 15 , SO 2 N(R 14 ) 2 , N(R 16 )C(O)R 15 , N(R 16 )SO 2 R 15 , N(R 16 )C(O)N(R 14 ) 2 , N(R 16 )SO 2 N(R 14 ) 2 , N(R 14 ) 2 , P(O)(R 15 ) 2 , CH 2 C(O)R 15 , CH 2 C(O)N(R 14 ) 2
  • R 18 is absent, wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, or heteroaryl in R 14 , R 15 , R 16 , or R 17 , is optionally further substituted, preferably R 17 is selected from C 1-6 alkyl, C 5-10 heteroaryl, C 4 -wheterocycloalkyl, N(R 14 ) 2 , N(R 16 )C(O)R 15 , N(R 16 )SO 2 R 15 , C(O)N(R 14 ) 2 , and SO 2 N(R 14 ) 2 , wherein said alkyl, alkenyl
  • R 17 is selected from H, F, NH 2 , and an optionally substituted C 5-10 heteroaryl or C 4-10 heterocycloalkyl, preferably R 17 is an optionally substituted C 5-10 heteroaryl or C 4-10 heterocycloalkyl.
  • R 17 is an optionally substituted C 4-10 heterocycloalkyl, wherein said heterocycloalkyl may be mono or bicyclic and include from 1 to 3 heteroatoms, preferably wherein X 7 is N.
  • the heterocycloalkyl is substituted, for instance, with at least one group selected from F, OH, oxo, CN, C 1-4 alkyl and OC 1-4 alkyl, wherein said C 1-4 alkyl is optionally further substituted (e.g. with F, OH, OCiwalkyl, etc.).
  • the heterocycloalkyl may be selected from optionally substituted piperidine, piperazine, thiomorpholine, and morpholine groups, or a bicyclic structure (bridged or spiro) containing a piperidine, piperazine, thiomorpholine, or morpholine ring.
  • X 7 is C, for instance, X 7 is C and R 18 is selected from Ciwalkyl, C 5- wheteroaryl, C 3-10 cycloalkyl, C 4-10 heterocycloalkyl, C(O)R 15 , C(O)N(R 14 ) 2 , SO 2 R 15 , SO 2 N(R 14 ) 2 , N(R 16 )C(O)R 15 , N(R 16 )SO 2 R 15 , N(R 16 )C(O)N(R 14 ) 2 , N(R 16 )SO 2 N(R 14 ) 2 , N(R 14 ) 2 , P(O)(R 15 ) 2 , CH 2 C(O)R 15 , CH 2 C(O)N(R 14 ) 2 , CH 2 SO 2 R 15 , CH 2 SO 2 N(R 14 ) 2 , CH 2 N(R 16 )C(O)R 15 ,
  • R 15 CH 2 N(R 16 )SO 2 R 15 , CH 2 N(R 16 )C(O)N(R 14 ) 2 , CH 2 N(R 16 )SO 2 N(R 14 ) 2 , and CH 2 N(R 14 ) 2 , wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, in R 14 , R 15 , R 16 , or R 18 , or heteroaryl is optionally further substituted, preferably R 18 is selected from C(O)N(R 14 ) 2 , SO 2 R 15 , and SO 2 N(R 14 ) 2 .
  • R 17 is selected from H, halo, OH, C 1-6 alkyl, N(R 14 ) 2 , and an optionally substituted C 5-10 heteroaryl.
  • R 17 is selected from H, F, NH 2 , and an optionally substituted C 5-10 heteroaryl, preferably H, F, or NH 2 .
  • R 14 is independently in each occurrence selected from H, optionally substituted C 1-6 alkyl, optionally substituted C 3-10 cycloalkyl, optionally substituted C 4- 10 heterocycloalkyl, and optionally substituted C 5-6 heteroaryl, or two R 14 are taken together with their adjacent nitrogen atom to form a C 4-10 heterocycloalkyl group.
  • R 17 is N(R 14 ) 2 wherein said R 14 are taken together with their adjacent nitrogen atom to form a C 4-10 heterocycloalkyl group, wherein said heterocycloalkyl may be mono or bicyclic and include from 1 to 3 heteroatoms, preferably wherein X 7 is N.
  • the heterocycloalkyl is substituted, for instance, with at least one group selected from F, OH, oxo, CN, C 1-4 alkyl and OC 1-4 alkyl, wherein said C 1-4 alkyl is optionally further substituted (e.g. with F, OH, OC 1-3 alkyl, etc.).
  • the heterocycloalkyl may be selected from optionally substituted piperidine, piperazine, thiomorpholine, and morpholine groups, or a bicyclic structure (bridged or spiro) containing a piperidine, piperazine, thiomorpholine, or morpholine ring.
  • R 1 is selected from: wherein R 14 , R 17 , and R 27 are as defined herein and ( — ) represents a bond serving as a point of attachment between R 1 and the rest of the molecule.
  • R 1 is selected from:
  • R 14 , R 17 , and R 27 are as defined herein and ( — ) represents a bond serving as a point of attachment between R 1 and the rest of the molecule.
  • exemplary R 1 groups are illustrated as A1 to A550 defined as follows:
  • ( — ) represents a bond serving as a point of attachment between R 1 and the rest of the molecule.
  • R 1 is selected from groups A1 to A550 or R 1 is selected from groups A1 to A3, A8, A19, A20, A22, A23, A25, A28, A29, A32 to A39, A60, A63 to A66, A69, A72 to A78, A81 to A83, A86, A89, A96, A100, A101 , A104, A105, A109 to A111 , A113, A115, A118, A121 to A123, A127 and A132, for instance, R 1 is selected from groups A1 to A3, A8, A19, A22, A25, A28, A29, A32, A36, A37, A64, A67, A74, A77, A78, A82, A83, A89, A96, A109, A110 and A111.
  • R 1 is selected from groups A1 to A3, A8, A19, A20, A22, A23, A25, A28, A29, A32 to A39, A60, A63 to A66, A69, A72 to A78, A81 to A83, A86, A89, A93, A96, A100, A101 , A104, A105, A109 to A111 , A113, A115, A118, A121 to A123, A127 and A132, for instance, R 1 is selected from groups A1 to A3, A8, A19, A22, A25, A28, A29, A32, A36, A37, A64, A67, A74, A77, A78, A82, A83, A89, A93, A96, A109, A110 and A111 .
  • Exemplary compounds as defined herein include each single compound covered in Tables 3 and 4 under Examples 1 to 159.
  • Examples of preferred compounds are, namely, Examples 2, 4, 6, 7, 14, 16, 18, 30, 31 , 33 to 37, 40, 43 to 46, 49, 51 to 60, 81 , 84 to 88, 90, 93 to 99, 102 to 105, 108, 111 , 112, 116 to 119, 122, 126, 127, 130, 131, 135 to 137, 139, 141 , 144, 147, 148, 149, 153, and 158, or a salt and/or solvate thereof.
  • Examples of more preferred compounds include Examples 4, 6, 7, 14, 16, 18, 30, 33, 35, 36, 37, 40, 43 to 45, 49, 51 , 56 to 58, 85, 88, 95, 98, 99, 103, 104, 105, 111 , 112, 116, 122, 135, and 136, or a salt and/or solvate thereof.
  • any of the above compounds may be in any amorphous, crystalline or polymorphic form, including any salt or solvate form, or a mixture thereof.
  • the compounds of the present description may be further modified by appending various functionalities via any synthetic means delineated herein to enhance selective biological properties. Such modifications are known in the art and include those which increase biological penetration into a given biological system (e.g., blood, lymphatic system, central nervous system), increase oral availability, increase solubility to allow administration by injection, alter metabolism and alter rate of excretion.
  • the term "effective amount” means that amount of a drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system, animal or human that is being sought, for instance, by a researcher or clinician.
  • therapeutically effective amount means any amount which, as compared to a corresponding subject who has not received such amount, results in treatment, healing, prevention, or amelioration of a disease, disorder, or symptom thereof, or a decrease in the rate of advancement of a disease or disorder.
  • the term also includes within its scope amounts effective to enhance normal physiological function.
  • treatment refers to reversing, alleviating, delaying the onset of, or inhibiting the progress of a disease or disorder, or one or more symptoms thereof, as described herein.
  • treatment may be administered after one or more symptoms have developed.
  • treatment may be administered in the absence of symptoms.
  • treatment may be administered to a susceptible individual prior to the onset of symptoms (e.g., in light of a history of symptoms and/or in light of genetic or other susceptibility factors). Treatment may also be continued after symptoms have resolved, for example to prevent or delay their recurrence.
  • the disease or condition to be treated is a proliferative disease or disorder or a kinase-mediated disease or disorder. More specifically, the disease or disorder to be treated include a proliferative disease or disorder, a developmental anomaly caused by dysregulation of the RAS-ERK signaling cascade (RASopathies), an inflammatory disease or an immune system disorder.
  • RASopathies a developmental anomaly caused by dysregulation of the RAS-ERK signaling cascade
  • the proliferative disease or disorder to be treated is a neoplasm, an inflammatory disease or condition or a developmental anomaly, involving a constitutively activating mutation in RAS and/or RAF genes (e.g. KRAS and/or ARAF, BRAF or CRAF mutations).
  • the disease or disorder may also be further associated with a receptor tyrosine kinase mutation or amplification (e.g. EGFR, HER2) or a mutation in a regulator of RAS downstream of the receptor (e.g. SOS1 gain of function, NF1 loss of function).
  • the compounds as defined herein are inhibitors of signal enzymes (ex.
  • B- and CRAF which are involved in controlling cell proliferation not only in tumors harboring RAF mutations (e.g. BRAF V600E ) but importantly also in the context of mutated RAS-driven cancers.
  • the present compounds may be used for example for the treatment of diseases connected with the activity of these signal enzymes and characterized by excessive or abnormal cell proliferation.
  • the disease or disorder is characterized by uncontrolled cell proliferation, i.e. a “proliferative disorder” or “proliferative disease”. More specifically, these diseases and disorders relate to cells having the capacity for autonomous growth, i.e. an abnormal state of condition characterized by rapidly proliferating cell growth which generally forms a distinct mass that show partial or total lack of structural organization and functional coordination with normal tissue.
  • the proliferative disorder or disease is defined as a “neoplasm”, “neoplastic disorder”, “neoplasia” “cancer,” and “tumor” which terms are collectively meant to encompass hematopoietic neoplasms (e.g. lymphomas or leukemias) as well as solid neoplasms (e.g. sarcomas or carcinomas), including all types of pre-cancerous and cancerous growths, or oncogenic processes, metastatic tissues or malignantly transformed cells, tissues, or organs, irrespective of histopathologic type or stage of invasiveness.
  • hematopoietic neoplasms e.g. lymphomas or leukemias
  • solid neoplasms e.g. sarcomas or carcinomas
  • Hematopoietic neoplasms are malignant tumors affecting hematopoietic structures (structures pertaining to the formation of blood cells) and components of the immune system, including leukemias (related to leukocytes (white blood cells) and their precursors in the blood and bone marrow) arising from myeloid, lymphoid or erythroid lineages, and lymphomas (related to lymphocytes).
  • Solid neoplasms include sarcomas, which are malignant neoplasms that originate from connective tissues such as muscle, cartilage, blood vessels, fibrous tissue, fat or bone.
  • Solid neoplasms also include carcinomas, which are malignant neoplasms arising from epithelial structures, including external epithelia (e.g., skin and linings of the gastrointestinal tract, lungs, and cervix), and internal epithelia that line various glands (e.g., breast, pancreas, thyroid).
  • carcinomas include leukemia, and hepatocellular cancers, sarcoma, vascular endothelial cancers, breast cancers, central nervous system cancers (e.g.
  • the disease or disorder is selected from colon cancer, lung cancer, pancreatic cancer, thyroid cancer, breast cancer and skin cancer.
  • neoplasm examples include melanoma, papillary thyroid carcinoma, colorectal, ovarian, breast cancer, endometrial cancer, liver cancer, sarcoma, stomach cancer, Barret's adenocarcinoma, glioma (including ependymoma), lung cancer (including non-small cell lung cancer), head and neck cancer, acute lymphoblastic leukemia, acute myelogenous leukemia, non-Hodgkin's lymphoma, and hairy-cell leukemia.
  • patients presenting one of the above-mentioned hematopoietic or solid neoplasms have previously received treatment with a RAS-ERK pathway-targeted inhibitor (including RTK, RAF, MEK or ERK inhibitor) but have developed resistance to the said inhibitor.
  • the inhibitor includes standard of care treatments such as vemurafenib, dabrafenib, cobimetinib, trametinib, YERVOY, OPDIVO or any combination of these pharmaceutical agents.
  • the disease to be treated is defined by developmental anomalies caused by dysregulation of the RAS-ERK signaling cascade (RASopathies: e.g. Noonan syndrome, Costello syndrome, LEOPARD syndrome, cardiofaciocutaneous syndrome and hypertrophic cardiomyopathy).
  • RASopathies e.g. Noonan syndrome, Costello syndrome, LEOPARD syndrome, cardiofaciocutaneous syndrome and hypertrophic cardiomyopathy.
  • the disease to be treated is defined as an inflammatory disease or immune system disorder.
  • inflammatory diseases or immune system disorders including inflammatory bowel disease, Crohn's disease, ulcerative colitis, systemic lupus erythematosis (SLE), rheumatoid arthritis, multiple sclerosis, thyroiditis, type 1 diabetes, sarcoidosis, psoriasis, allergic rhinitis, asthma, COPD (chronic obstructive pulmonary disease).
  • the compounds as herein defined are inhibitors of RAS-ERK signaling and cellular proliferation in tumor cells bearing at least one mutated RAS or RAF genotype, without or substantially without inducing the paradoxical pathway.
  • patient or subject refers to an animal such as a mammal.
  • a subject may therefore refer to, for example, mice, rats, dogs, cats, horses, cows, pigs, guinea pigs, primates including humans and the like.
  • the subject is a human.
  • the present description therefore further relates to a method of treating a subject, such as a human subject, suffering from a proliferative disease or disorder, e.g. a RAF-mutated and/or mutated RAS-driven cancer.
  • the method comprises administering a therapeutically effective amount of a compound as defined herein, to a subject in need of such treatment.
  • the present description provides a method of treating a disorder (as described herein) in a subject, comprising administering to the subject identified as in need thereof, a compound of the present description.
  • a disorder as described herein
  • the identification of those patients who are in need of treatment for the disorders described above is well within the ability and knowledge of one skilled in the art.
  • Certain of the methods for identification of patients which are at risk of developing the above disorders which can be treated by the subject method are appreciated in the medical arts, such as family history, and the presence of risk factors associated with the development of that disease state in the subject patient.
  • a clinician skilled in the art can readily identify such candidate patients, by the use of, for example, clinical tests, physical examination, medical/family history, and genetic determination.
  • a method of assessing the efficacy of a treatment in a subject includes determining the pretreatment symptoms of a disorder by methods well known in the art and then administering a therapeutically effective amount of a compound of the present description, to the subject. After an appropriate period of time following the administration of the compound (e.g., 1 week, 2 weeks, one month, six months), the symptoms of the disorder are determined again.
  • the modulation (e.g., decrease) of symptoms and/or of a biomarker (e.g. pERK or pMEK) of the disorder indicates efficacy of the treatment.
  • the symptoms and/or biomarker of the disorder may be determined periodically throughout treatment. For example, the symptoms and/or biomarker of the disorder may be checked every few days, weeks or months to assess the further efficacy of the treatment.
  • a decrease in symptoms and/or biomarker of the disorder indicates that the treatment is efficacious.
  • the therapeutically effective amount of a compound as defined herein can be administered to a patient alone or in a composition, admixed with a pharmaceutically acceptable carrier, adjuvant, or vehicle.
  • compositions of this disclosure include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose- based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxy
  • compositions described herein may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, or via an implanted reservoir.
  • parenteral as used herein includes subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. Other modes of administration also include intradermal or transdermal administration.
  • Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs.
  • the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.
  • the oral compositions can also include adjuvant
  • sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents.
  • the sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1 ,3-butanediol.
  • acceptable vehicles and solvents that may be employed are water, Ringer's solution, ll.S.P. and isotonic sodium chloride solution.
  • sterile, fixed oils are conventionally employed as a solvent or suspending medium.
  • any bland fixed oil can be employed including synthetic mono- or diglycerides.
  • fatty acids such as oleic acid are used in the preparation of injectables.
  • Injectable formulations can be sterilized, for example, by filtration through a bacterial -retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.
  • biodegradable polymers examples include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the compound in liposomes or microemulsions that are compatible with body tissues.
  • compositions for rectal administration are preferably suppositories which can be prepared by mixing the compounds of the present description with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum and release the active compound.
  • suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum and release the active compound.
  • Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.
  • the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone (PVP), sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and
  • Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.
  • the solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.
  • the composition can also be in micro-encapsulated form with one or more excipients as noted above.
  • the solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings and other coatings well known in the pharmaceutical formulating art.
  • the active compound may be admixed with at least one inert diluent such as sucrose, lactose or starch.
  • Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose.
  • the dosage forms may also comprise buffering agents. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner.
  • buffering agents include polymeric substances and waxes.
  • Dosage forms for topical or transdermal administration of a compound of the present description include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches.
  • the active component is admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers as may be required.
  • Ophthalmic formulation, ear drops, and eye drops are also contemplated as being within the scope of the present description.
  • the description contemplates the use of transdermal patches, which have the added advantage of providing controlled delivery of a compound to the body.
  • Such dosage forms can be made by dissolving or dispensing the compound in the proper medium.
  • Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.
  • compositions provided herein may also be administered by nasal aerosol or inhalation.
  • Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promotors to enhance bioavailability, fluorocarbons, and/or other conventional solubilizing or dispersing agents.
  • compositions provided herein may be formulated for oral administration. Such formulations may be administered with or without food. In some embodiments, pharmaceutically acceptable compositions of this disclosure are administered without food. In other embodiments, pharmaceutically acceptable compositions of this disclosure are administered with food.
  • compositions may be formulated such that a dosage of between 0.01 - 100 mg/kg body weight/day of the inhibitor can be administered to a patient receiving these compositions.
  • a specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, the judgment of the treating physician, and the severity of the symptoms associated with the proliferative disease or disorder.
  • the amount of a provided compound in the composition will also depend upon the particular compound in the composition.
  • Compounds or compositions described herein may be administered using any amount and any route of administration effective for treating or lessening the severity of the symptoms as contemplated herein.
  • the exact amount required will vary from subject to subject, depending on the species, age, and general condition of the subject, the severity of the infection, the particular agent, its mode of administration, and the like.
  • Provided compounds are preferably formulated in unit dosage form for ease of administration and uniformity of dosage.
  • the expression "unit dosage form” as used herein refers to a physically discrete unit of agent appropriate for the patient to be treated. It will be understood, however, that the total daily usage of the compounds and compositions of the present disclosure will be decided by the attending physician within the scope of sound medical judgment.
  • compositions of this disclosure can be administered to humans and other animals orally, rectally, parenterally, intracisternally, intraperitoneally, topically (as by powders, ointments, or drops), buccally, as an oral or nasal spray, or the like, depending on the severity of the infection being treated.
  • provided compounds may be administered orally or parenterally at dosage levels of about 0.01 mg/kg to about 50 mg/kg and preferably from about 1 mg/kg to about 25 mg/kg, of subject body weight per day, one or more times a day, to obtain the desired therapeutic effect.
  • the total daily inhibitory dose of the compound of the present description administered to a subject in single or in divided doses can be in amounts, for example, from 0.01 to 50 mg/kg body weight or more usually from 0.1 to 25 mg/kg body weight.
  • Single dose compositions may contain such amounts or submultiples thereof to make up the daily dose.
  • treatment regimens according to the present description comprise administration to a patient in need of such treatment from about 10 mg to about 1000 mg of the compound(s) of the present description per day in single or multiple doses.
  • additional therapeutic agents may also be present in the compositions of this disclosure or administered separately as part of a dosage regimen, e.g. an additional chemotherapeutic agent.
  • additional therapeutic agents include antiproliferative compounds such as aromatase inhibitors; anti-estrogens; anti-androgens; gonadorelin agonists; topoisomerase I inhibitors; topoisomerase II inhibitors; microtubule active agents; alkylating agents; retinoids, carotenoids, tocopherol; cyclooxygenase inhibitors; MMP inhibitors; antimetabolites; platin compounds; methionine aminopeptidase inhibitors; bisphosphonates; antiproliferative antibodies; heparanase inhibitors; inhibitor of Ras oncogenic isoforms; telomerase inhibitors; proteasome inhibitors; compounds used in the treatment of hematologic malignancies; kinesin spin
  • the treatment may also be complemented with other treatments or interventions such as surgery, radiotherapy (e.g., gamma-radiation, neutron beam radiotherapy, electron beam radiotherapy, proton therapy, brachytherapy, and systemic radioactive isotopes), a biologic response modifier (e.g., an interferon, an interleukin, tumor necrosis factor (TNF), and agents used to attenuate an adverse effect.
  • radiotherapy e.g., gamma-radiation, neutron beam radiotherapy, electron beam radiotherapy, proton therapy, brachytherapy, and systemic radioactive isotopes
  • a biologic response modifier e.g., an interferon, an interleukin, tumor necrosis factor (TNF)
  • agents used to attenuate an adverse effect e.g., gamma-radiation, neutron beam radiotherapy, electron beam radiotherapy, proton therapy, brachytherapy, and systemic radioactive isotopes
  • an interferon e.g.
  • BSA bovine serum albumin
  • CDCI 3 deuterated chloroform
  • DIEA N,N-diisopropylethylamine (Huenig’s base)
  • EDTA ethylenediamine tetraacetic acid
  • FBS fetal bovine serum
  • HATU O-(7-azabenzotriazol-1-yl)-N,N,N’,N’,-tetramethyluronium hexafluorophosphate
  • PBS phosphate buffered saline
  • pERK phosphorylated extracellular signal-regulated kinase
  • TBST Tris buffered saline with 0.2% Tween-20
  • TEV tobacco etch virus protease
  • Ts para-Toluenesulfonate
  • Y MIN minimal data point of a dosage-activity curve
  • Preparative HPLC was performed using an Agilent instrument using a Phenomenex-Kinetex C18, (21x100mm, 5 pm) column at a flow rate of 20 mL/min (RT) and UV detection at 220 and 254 nm.
  • the mobile phase consisted of Solvent A (5% MeOH, 95% water + 0.1% formic acid) and Solvent B (95% MeOH, 5% water + 0.1% formic acid) unless stated otherwise.
  • 0.05% TFA or 0.1 % AcOH or other additives were occasionally used as additives instead of 0.1 % formic acid in both solvents.
  • MeCN was also used instead of MeOH in both mobile phases for more challenging separations as specified in the text. Specific gradient conditions are provided in the examples but the following is representative: T(0) ⁇ T(3 min) isocratic using between 10 to 50% solvent B depending on compound polarity, followed by a 12 minutes gradient to 100% solvent B. Last 5 minutes 100% solvent B.
  • LCMS analyses were performed on an Agilent instrument. Liquid chromatography was performed on a Phenomenex Kinetex C18 column (2.6 pm; 100 A; 3 X 30 mm) at a flow rate of 1.5 mL/min (RT) with UV detection at 220 and 254 nm.
  • the mobile phase consisted of solvent A (95% H 2 O I 5% MeOH I 0.1% AcOH) and solvent B (95% MeOH I 5% H 2 O I 0.1% AcOH) using the following gradient: T(0) 100% A ⁇ T(0.5 min) 100% B ⁇ isocratic 100% B to T(2 min). MS detection was performed in parallel using APCI detection in both positive and negative modes.
  • 2,6-difluoroaniline A-1 can be converted to its acetanilide A-2 using an acetylating agent such as acetic anhydride and converted to mono-protected dianiline A-3 as described in WO 2012/101238A1.
  • Sulfonylation to sulfonamide A-4 can be achieved using sulfonylating reagents such as sulfonyl chlorides in the presence of an organic base such as pyridine, with or without a catalyst such as 4-dimethylaminopyridine and solvents such as dichloromethane or tetrahydrofuran.
  • Treatment of acetanilide A-4 with aqueous hydrochloric acid in the presence of a co-solvent such as an alcohol provides aniline salt A-5.
  • 2-amino-2-cyanoacetamide A-6 can be transformed into 5-amino-2- (methylthio)thiazole-4-carboxamide A-8 following a 2 step procedure described in Heterocycl. Commun., 2014, 20, 175.
  • Intermediate A-8 can then be cyclized to the corresponding pyrimidone by heating in formamide as described in Indian J. Chem., 2010, 49B, 1229.
  • chlorinating agents such as thionyl chloride or phosphoryl chloride in the presence of a catalytic amount of DMF provides chloro compound A-10 following a procedure which is also described in Indian J. Chem., 2010, 49B, 1229.
  • Intermediates A-11 can be obtained by heating chloropyrimidine A-10 with aniline salts A-5 in an organic acid such as acetic acid.
  • Inhibitors of general formulae W-l are prepared from intermediates of general formulae A-11 by a two-step procedure involving first oxidation of the thiomethyl group generally to a mixture of the corresponding methylsulfoxide and methylsulfone which is then reacted with a nucleophile (e.g. a 1° or a 2° amine or a NH-containing heterocycle, etc.). The latter step is usually carried out in the presence of a base (e.g.
  • an organic base such as DIEA, trimethylamine, pyridine and the like or an inorganic base such as potassium carbonate, cesium carbonate and sodium carbonate
  • a solvent such as DMSO or NMP at a temperature ranging from 70 to 140 °C.
  • Inhibitors of general formulae W-ll are prepared as described in Synthetic Method B.
  • an organic or inorganic base such as CS 2 CO 3 , KOtBu, DIEA, trimethylamine, pyridine and the like
  • a solvent such as THF, DMSO or NMP
  • inorganic base e.g. NaOH or KOH
  • miscible organic solvent e.g. methanol, ethanol, THF, dioxane and the likes
  • aqueous NH 4 CI or KHSO 4 or organic acid (e.g. aqueous citric or acetic acid) provided the corresponding carboxylic acid intermediates B-1 or B-2.
  • Coupling of intermediates B-1 or B-2 with amines using standard amide coupling reagents e.g. TBTU, HATU, DCC, EDC and the likes
  • amide derivatives of general formulae W-ll amide derivatives of general formulae W-ll.
  • Cross-coupling of commercially available bromobenzimidazole C-1 to heteroaryl boronic acids or boronate esters can be performed under palladium-catalyzed Suzuki-Miyaura cross-coupling conditions in the presence of a base such as sodium or potassium carbonate in a solvent such as dioxane or dimethoxyethane and water, to provide intermediates C-2.
  • a base such as sodium or potassium carbonate
  • a solvent such as dioxane or dimethoxyethane and water
  • Substituted benzimidazole derivatives C-2 can then be attached to intermediates A-11 following the oxidation of the methylsulfide moiety as described in the general protocol of Method A, to provide inhibitors of general structure W-lll.
  • Unprotected 3-indolesulfonyl chloride D-2 can be prepared as described in Org. Lett. 2011 , 13, 3588. This reagent can then be converted to the corresponding sulfonamide D-3 by reacting with a 1° or a 2° amine in the presence of an organic base such as DIEA or triethylamine. D-3 fragments can then be attached to intermediates A-11 following the oxidation of the methylsulfide moiety as described in the general protocol of Method A, to provide inhibitors of general structure W-IV.
  • inhibitors of general structure W-IV can be accessed through N-tosyl protected indole-3-sulfonyl chloride E-1 , prepared following the procedure described in Chemical and Pharmaceutical Bulletin 2009, 57, 591.
  • E-1 can then be converted to the corresponding sulfonamide E-2 by reacting with a 1 ° or a 2° amine in the presence of an organic base such as DIEA or triethylamine followed by removal of the tosyl protecting group with an aqueous inorganic base such as KOH.
  • E-2 fragments can then be attached to intermediates A-11 following the oxidation of the methylsulfide moiety as described previously.
  • 3-lndolethiocyanate F-1 (prepared following the procedure described in Phosphorus, Sulfur and Silicon and the Related Elements 2014, 189, 1378) is reduced to the corresponding sulfide salt using a reducing agent such as sodium sulfide nonahydrate and directly alkylated without isolation with an alkyl halide to provide sulfide intermediates F-2.
  • Sulfide intermediates F-2 are then converted to sulfone intermediates F-3 using an oxidizing agent such as 3-chloroperoxybenzoic acid.
  • Final inhibitors of the general structure W-V are then obtained by attaching indolesulfones F-3 with A-11 intermediates following the oxidation of the methylsulfide moiety as described previously.
  • a bright red solution of commercially available 3-fluoro-2-nitroaniline G-1 reacts with primary or secondary amines in solvents such as MeCN, DMSO, NMP or DMF and in the presence of an inorganic base such as potassium carbonate or an organic base such as DIEA to provide intermediates G-2 upon heating at temperature ranging from 40 °C to 120 °C under thermal or microwave conditions.
  • solvents such as MeCN, DMSO, NMP or DMF
  • an inorganic base such as potassium carbonate or an organic base such as DIEA
  • Sulfonyl Chlorides The following sulfonyl chlorides were obtained from commercial sources and used as received: 2-chlorobenzenesulfonyl chloride, 2,3-dichlorobenzenesulfonyl chloride, 3-chloro-2- methylbenzenesulfonyl chloride, 3-fluoro-2-methylbenzenesulfonyl chloride.
  • Step 2 To a solution of 2-fluoro-3-methylanisole from step 1 (1.00 g, 7.13 mmol) in DCM (5.6 mL) was added a solution of chlorosulfonic acid (1.13 mL, 16.5 mmol) in DCM (5.6 mL) over a 5 minute period. The pale brown reaction mixture containing a viscous liquid layer was stirred at RT for 10 min and then quenched by pouring into a mixture of water (10 mL) and ice (5 g).
  • Acetanilide A-2 can be prepared by acetylation of 2,6- difluoroaniline A-1 with acetic anhydride following the literature procedure described in Bioorg. Med. Chem. 2016, 24, 2215. Intermediate A-2 was converted to acetanilide A-3 by sequential nitration followed by reduction of the nitro group to the aniline as described in WO 2012/101238A1.
  • Steps 1 and 2 Preparation of intermediate A-8: These two steps were performed as described by Wang et al. in Heterocycl. Commun., 2014, 20, 175.
  • Step 3 Preparation of intermediate A-9 (adapted from Indian J. Chem., 2010, 49B, 1229): 5- amino-2-methylsulfanyl-thiazole-4-carboxamide (3.19 g, 16.855 mmol) was divided equally into two microwaveable 20 mL vials to which was added formamide (13.4 mL, 337 mmol) in each. The vials were sealed and heated in the microwave at 185 °C each for 20 minutes then at 190 °C for another 15 minutes.
  • Step 4 Preparation of intermediate A-10 (adapted from Indian J. Chem., 2010, 49B, 1229): 2- methylsulfanyl-6H-thiazolo[5,4-d]pyrimidin-7-one (1.00 g, 5.02 mmol) was suspended in thionyl chloride (10 mL, 137 mmol) then 3 drops of DMF were added. The resulting mixture was brought to reflux in an oil bath set at 90 °C. After 3 hours of stirring, the mixture had become a clear solution. It was allowed to cool to room temperature then diluted with some toluene and concentrated to dryness under reduced pressure.
  • Example 4 2,3- dichloro-N-[2,4-difluoro-3-[(2-methylsulfonylthiazolo[5,4-d]pyrimidin-7- yl)amino]phenyl]benzenesulfonamide (50 mg, 0.0883 mmol) and 1-H-benzimidazole (26 mg, 0.221 mmol) were dissolved in 0.6 mL of NMP in a 1 dram vial. N,N-diisopropylethylamine (0.077 mL, 0.441 mmol) was then added and the mixture was heated with stirring at 105 °C for 22 hours.
  • the reaction was allowed to cool to room temperature then quenched by the addition of a formic acid (0.1 mL) solution in methanol (1 mL).
  • the solution was then filtered and purified by reversed- phase preparative HPLC using a 40% to 90% MeOH in water gradient with 0.1 % formic acid modifier. The appropriate fractions were pooled and concentrated. The residues were lyophilized from a water and MeCN mixture.
  • Steps 2 and 3 Preparation of carboxylic acid B-1 : The crude mixture of sulfone and sulfoxide from above (300 mg, 0.567 mmol) and indole-3-carboxylic acid methyl ester (149 mg, 0.850 mmol) were charged in a 20 mL vial and suspended in NMP (3.6 mL). DIPEA (0.49 mL, 2.83 mmol) was then added to the orange solution and the reaction mixture was stirred at 105 °C for 24h. The reaction mixture was allowed to cool to room temperature and a 4N solution of sodium hydroxide (0.85 mL, 3.40 mmol) was added. The resulting mixture was stirred at 50 °C for another hour.
  • Step 4 Preparation of W-ll
  • Example 28 1-[7-[2,6-difluoro-3-[(3-fluoro-2-methyl- phenyl)sulfonylamino]anilino]thiazolo[5,4-d]pyrimidin-2-yl]indole-3-carboxylic acid (35 mg, 0.0573 mmol) and HATLI (44 mg, 0.115 mmol) were dissolved in NMP (1 mL) followed by DI PEA (0.060 mL, 0.344 mmol). The orange solution was allowed to stir at room temperature for 2-3 minutes and then N-methyl-2-(pyridin-2-yl)ethan-1-amine (0.016 mL, 0.115 mmol) was added.
  • reaction mixture was allowed to stir at room temperature for 2 hours.
  • the reaction was then quenched by the addition of 0.5 mL of formic acid, diluted with a bit of DMSO and then purified by preparative reversed-phase HPLC to provide the compound of example 28 (9.8 mg, 23 % yield) as a beige solid after lyophilisation from a MeCN/water mixture.
  • Step 1 Preparation of the methylsulfone and methylsulfoxide mixture from intermediate A-11 was performed as described above for Example 28 (step 1).
  • Steps 2 and 3 Preparation of carboxylic acid B-2: The crude mixture of sulfone and sulfoxide from above (300 mg, 0.567 mmol) and methyl indazolyl-3-carboxylate (150 mg, 0.850 mmol) were charged in a 20 mL vial and suspended in NMP (3.6 mL). DIPEA (0.49 mL, 2.83 mmol) was then added to the orange solution and the reaction mixture was stirred at 105 °C for 24h. It was then was allowed to cool to room temperature and 4N sodium hydroxide (0.85 mL, 3.40 mmol) in water was added. The resulting mixture was stirred at 50 °C for another hour.
  • Step 4 Preparation of W-ll
  • Example 23 1-[7-[2,6-difluoro-3-[(3-fluoro-2-methyl- phenyl)sulfonylamino]anilino]thiazolo[5,4-d]pyrimidin-2-yl]indazole-3-carboxylic acid (35 mg, 0.0572 mmol) and HATU (44 mg, 0.114 mmol) were dissolved in NMP (1 mL). DIPEA (0.060 mL, 0.343 mmol) was then added.
  • Step 1 Preparation of substituted benzimidazole intermediate C-2: Bromobenzimidazole C-1 (70 mg, 0.355 mmol), potassium carbonate (196 mg, 1.42 mmol) and 3-pyridylboronic acid (57 mg, 0.46 mmol) were charged in a 4 mL vial and dioxane (2 mL) and water (0.7 mL) were added. Argon gas was bubbled through the mixture for 1 minute and then tetrakis(triphenylphosphine)palladium (0) (16.4 mg, 0.014 mmol) was added.
  • Step 2 Preparation of the methylsulfone and methylsulfoxide mixture from intermediate A-11 was performed as described above in Method B for Example 28 (step 1).
  • Step 2 The crude pyridinium 1 H-indole-3-sulfonate from step 1 (4.60 g, 16.7 mmol) was dissolved in a 1 :1 mixture of sulfolane:acetonitrile (50 mL). The white suspension was cooled to 0 °C, and POCI 3 (3.42 mL, 36.7 mmol) was added dropwise under stirring resulting a light brown solution. The reaction was heated to 70 °C for 1 h. After 1 h, the orange solution was cooled to 0 °C. The cold orange solution was added dropwise to 250 mL of iced water. During the addition, a white precipitate was formed. The solid was filtered, washed with water and dried under vacuum to afford 1 H-indole-3-sulfonyl chloride D-2 (740 mg, 21 % yield) as a grey solid.
  • Step 3 1 H-indole-3-sulfonyl chloride (0.40 g, 1.86 mmol) was charged into a 25 mL flask to which was added 6 mL of THF. The solution was cooled to 0 °C then 1 -methylpiperazine (0.41 mL, 3.71 mmol) was added dropwise. The mixture went from a red solution to a pale yellow solution with a large amount of gummy solid in the bottom of the flask. DI PEA (0.97 mL, 5.56 mmol) was then added and the mixture was allowed to warm to room temperature. After 30 minutes of stirring at room temperature, the mixture was diluted with EtOAc and water.
  • Step 4 Preparation of the methylsulfone and methylsulfoxide mixture from intermediate A-11 was performed as described above in Method B for Example 28 (step 1).
  • Step 1 Preparation of 3-indolesulfonamide fragment E-2: To a vial with a stirring bar was added 1-(p-tolylsulfonyl)indole-3-sulfonyl chloride (117 mg, 0.316 mmol) (Chemical and Pharmaceutical Bulletin 2009, 57, 591) in anhydrous THF (1.5mL). The solution was cooled to 0 °C and DIEA (0.11 mL, 0.633 mmol) was added dropwise. N-(2-methoxyethyl)ethylamine (0.039 mL, 0.316 mmol) was then added dropwise and the reaction mixture was gently warmed to room temperature. The reaction was stirred at room temperature for 1h.
  • Step 2 Preparation of the methylsulfone and methylsulfoxide mixture from intermediate A-11 was performed as described above in Method B for Example 28 (step 1).
  • step 7 in general method A.
  • 1 H-indol-3-yl- thiocyanate Phosphorus, Sulfur and Silicon and the Related Elements 2014, 189, 1378)
  • Step 3 Preparation of the methylsulfone and methylsulfoxide mixture from intermediate A-11 was performed as described above in Method B for Example 28 (step 1).
  • Step 1 4-Methylpiperidin-4-ol (0.24 g, 1.84 mmol) and potassium carbonate (0.49 g, 3.52 mmol) were added to a solution of 3-fluoro-2-nitro-aniline (0.25 g, 1.60 mmol) in MeCN (2.6 mL) . The resulting mixture was stirred at 85 °C for 10 h. MeCN was removed under reduce pressure and EtOAc was added. The suspension was centrifuged and the supernatant poured into a flask. The solution was concentrated and the crude 1-(3-amino-2-nitro-phenyl)-4-methyl- piperidin-4-ol (0.40 g, 94% yield) was used without further purification for the next step. MS m/z 252.2 (MIT).
  • Step 2 Iron (0.37 g, 6.70 mmol) and ammonium chloride (0.36 g, 6.70 mmol) were added to a mixture of 1-(3-amino-2-nitro-phenyl)-4-methyl-piperidin-4-ol 0.34 g, 1.34 mmol) in iPrOH (6.5 mL) and formic aicd (1.9 mL, 49.6 mmol). The resulting mixture was heated to 90 °C and stirred for 10h. The reaction mixture was cooled down to room temperature and filtered through Celite®.
  • Step 1 Preparation of 4-(trifluoromethyl)-1H-benzo[d]imidazole (for Example 17): In a 10mL vial was added 3-(trifluoromethyl)benzene-1,2-diamine (280 mg, 1.59 mmol) in formic acid (2.0 mL). The reaction was heated to 100 °C and left to stir for 3 hours. The reaction was cooled to room temperature and the formic acid was removed under reduced pressure. The residue was taken up in a saturated solution of NaHCO 3 affording a precipitate which was homogenized by sonication. The solids were collected by filtration, washed with water, briefly dried on the filter and collected.
  • Step 1 Preparation of 4,5-difluoro-1 H-benzimidazole (for Example 49): 2,3-difluoro-6-nitro-aniline (800 mg, 4.60 mmol) was charged in a flask along with ammonium chloride (2.46 g, 46.0 mmol) and iron powder (2.57 g, 46.0 mmol). I PA (11.5 mL) was then added followed by formic acid (12 mL, 305 mmol). The flask was equipped with a reflux condenser and the reaction flask was immersed in an oil bath set to 80 °C and stirred for 5 hours.
  • Step 1 Preparation of 3-methylsulfonyl-1 H-indole: Indole (182 mg, 1.55 mmol) was dissolved in THF (17mL) under nitrogen at room temperature. Potassium tert-butoxide (192 mg, 1.71 mmol) was added and the mixture stirred 30 min. Triethylborane (0.22 mL, 1.55 mmol) was added dropwise at RT and stirred for 30 min. Methanesulfonyl chloride (0.13 mL, 1.71 mmol) was added and the mixture was stirred at -15 °C 5 days. A saturated aqueous solution of NaHCO 3 was added then the mixture was extracted with EtOAc.
  • Step 1 Preparation of 2-fluoro-3-(2-methoxyethoxy)-6-nitro-aniline (for Example 79): 2,3-difluoro- 6-nitro-aniline (800 mg, 4.60 mmol) was charged in a 25 mL flask and dissolved in 10 mL of DMF at room temperature affording a bright yellow solution. 2-methoxyethanol (2.2 mL, 27.6 mmol) was then added followed by potassium carbonate (2.54 g, 18.4 mmol). The color of the solution changed to bright red. The mixture was heated in an oil bath at 80 °C for 22 hours. After cooling, the mixture was diluted with EtOAc and washed with a saturated aqueous ammonium chloride solution.
  • Step 2 Preparation of the substituted benzimidazoles from the 2-nitroanilines described in step 1 was performed using the “one-pot” procedure described in Method G for Example 113 (step 2).
  • Step 1 Preparation of (5-chloro-1 H-indol-3-yl)-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)methanone: 5-chloro-1 H-indole-3-carboxylic acid (50 mg, 0.256 mmol) was dissolved in NMP (1.5 mL). 2-(1 H- Benzotriazole-1-yl)-1 ,1 ,3,3,-tetramethyluronium tetrafluoroborate (98 mg, 0.31 mmol) was added at room temperature followed by 3-Oxa-8-aza-bicyclo[3.2.1]octane HCI (42 mg, 0.28 mmol).
  • N,N- Diisopropylethylamine (0.18 mL, 1.02 mmol) was finally added and the mixture was allowed to stir for 1 hour. It was then partitioned between EtOAc and a saturated aqueous solution of ammonium chloride. The layers were separated and the aqueous layer was further extracted twice with EtOAc. The combined organic layers were washed with brine then dried over MgSO 4 , filtered and concentrated. The resulting residue was purified by silica gel chromatography using a 1 :1 EtOAc in DCM to 100% EtOAc gradient followed by 2% I PA in EtOAc.
  • Step 1 Preparation of 1-(1 H-indol-3-yl)propan-1-one (for Example 91): To a 100mL round-bottom flask, was added indole (200 mg, 1.71 mmol) in 7mL of DCM. The solution was cooled to 0 °C then a 1.8M (25% wt) solution of Et 2 AICI in toluene (1.42 mL, 2.56 mmol) was added dropwise. The reaction was left to stir for 30 min at 0 °C. Propionyl chloride in 6mL of DCM was added dropwise. The reaction was warmed to room temperature and left to stir for 2.5 hours.
  • the reaction was quenched with 3 eq. of NaOAc in 5mL of water.
  • the reaction was diluted with DCM and filtered through Celite® to remove the aluminum salts.
  • the filtrate was transferred to a separatory funnel and then water was added. 1M NaOH was added to help solubilize any remaining aluminum salts.
  • the layers were separated, and the organic layer was washed with water then brine. The organic layer was then dried over MgSO 4 filtered and then concentrated under reduced pressure.
  • the crude material was purified by silica gel chromatography using a 100% hexanes to 75% EtOAc in hexanes gradient.
  • Step 1 tert-Butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate (0.70 g, 3.30 mmol) and potassium carbonate (1.14 g, 8.24 mmol) were charged in a 25 mL flask and suspended in 10 mL of MeCN. benzyl 2-bromoethyl ether (0.57 mL, 3.63 mmol) was then added dropwise at room temperature. The resulting mixture was allowed to stir at room temperature for 22 hours then at 40 °C for 4 hours. The reaction mixture was diluted with EtOAc. The solids were removed by filtration and the filtrate was concentrated to dryness.
  • Step 2 tert-butyl 3-(2-benzyloxyethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (673 mg, 1.94 mmol) was charged in a 100 mL flask and dissolved in 5 mL of dioxane. A 4N solution of HCI in dioxane (1.94 mL, 7.76 mmol) was then added dropwise. After 1 hour of stirring, another portion of the HCI solution was added (2.91 mL, 11.64 mmol) and the mixture was warmed to 40 °C. After another 2 hours of stirring at that temperature, a gum had separated at the bottom of the flask.
  • Step 3 To a solution of 3-(2-benzyloxyethyl)-3,8-diazabicyclo[3.2.1]octane;dihydrochloride (615 mg, 1.93 mmol) in a 100 mL flask in 15 mL of methanol was added 10% palladium on charcoal (205 mg, 0.193 mmol). The flask was placed under reduced pressure then filled with hydrogen. This operation was repeated 3 more times then the mixture was stirred vigorously under a balloon atmosphere of hydrogen over weekend. The flask was flushed with nitrogen and the reaction mixture was filtered through a short pad of Celite®. The filtrate was concentrated and dried under reduced pressure.
  • Step 2 Preparation of 2-[(6-bromo-4-fluoro-benzimidazol-1-yl)methoxy]ethyl-trimethyl-silane and 2-[(5-bromo-7-fluoro-benzimidazol-1-yl)methoxy]ethyl-trimethyl-silane: 6-bromo-4-fluoro-1 H- benzimidazole (500 mg, 2.05 mmol) was dissolved in 10 mL of DMF then potassium carbonate (848 mg, 6.14 mmol) was added followed by SEM-CI (0.44 mL, 2.46 mmol) at room temperature. The mixture was allowed to stir at the same temperature for 18 hours.
  • Step 3 Preparation of 2-[7-fluoro-3-(2-trimethylsilylethoxymethyl)benzimidazol-5-yl]oxyethanol: The 3:1 mixture of isomers 2-[(6-bromo-4-fluoro-benzimidazol-1-yl)methoxy]ethyl-trimethyl-silane and 2-[(5-bromo-7-fluoro-benzimidazol-1-yl)methoxy]ethyl-trimethyl-silane (630 mg, 1.82 mmol) described above was charged in a 20 mL vial followed by 1 ,10-phenanthroline (33 mg, 0.18 mmol), copper (I) iodide (35 mg, 0.18 mmol) and cesium carbonate (1.19 g, 3.65 mmol).
  • Step 4 Preparation of 2-[(7-fluoro-3H-benzimidazol-5-yl)oxy]ethanol: 2-[7-fluoro-3-(2- trimethylsilylethoxymethyl)benzimidazol-5-yl]oxyethanol (308 mg, 0.94 mmol) was dissolved in 5 mL of THF in a 20 mL vial then a 1M THF solution of tetra-N-butylammonium fluoride (4.72 mL, 4.72 mmol) was added at room temperature. The vial was then sealed and the mixture was heated at 65 °C for 23 hours. The mixture was diluted with EtOAc and water with a bit of NaHCO 3 .
  • Step 1 To a 20mL vial was added 1H-benzimidazole-4-carboxylic acid (1.00 g, 6.17 mmol) in MeOH (6.2 mL). Sulfuric acid (723 ⁇ L, 13.6 mmol) was then added. The reaction was heated in an oil bath at 70 °C for 24 hours. It was allowed to cool to room temperature. The solvent was removed under reduced pressure. The crude material was taken up in a saturated aqueous solution of NaHCO 3 and extracted three times with EtOAc.
  • Step 2 To a 250 mL round-bottom flask was added methyl 1H-benzimidazole-4-carboxylate (750 mg, 4.26 mmol) from the previous step and 2-(trimethylsilyl)ethoxymethyl chloride, stabilized, tech. (982 ⁇ L, 5.53 mmol) in anhydrous THF (21 mL) under a N2 atmosphere.
  • Step 3 In a 20mL vial was added [1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]methanol (200 mg, 0.72 mmol) from the previous step and DIPEA (500 ⁇ L, 2.87 mmol) in MeCN (8 mL). methanesulfonyl chloride (167 ⁇ L, 2.16 mmol) was then added. The reaction was allowed to stir at room temperature overnight. The reaction was quenched with the addition of a saturated aqueous solution of NaHCO 3 and then extracted with EtOAc (x3). The combined organic layers were dried over MgSO 4 , filtered and then concentrated under reduced pressure.
  • the crude material was dissolved in DMF (8 mL) and then potassium cyanide (187 mg, 2.87 mmol) was added. The reaction was allowed to stir at room temperature overnight again. The reaction was quenched with the addition of a 10% aqueous solution of LiCI and then extracted with EtOAc (x3). The combined organic layers were dried over MgSO 4 , filtered and then concentrated under reduced pressure. The crude material was adsorbed onto Celite® and then purified by normal-phase silica gel chromatography using a 0% to 100% EtOAc in hexanes gradient.
  • Step 4 Sodium hydride (60 mg, 1.50 mmol) was added to 4 mL of DMF in a 20 mL vial. The mixture was cooled to 0 °C and then a solution of 2-[1-(2-trimethylsilylethoxymethyl)benzimidazol- 4-yl]acetonitrile (177 mg, 0.616 mmol) from the previous step in 4 mL of DMF was slowly added. Upon addition, the reaction turned an orange colour. The reaction was warmed to room temperature. After ⁇ 30min, the reaction turned a darker orange colour. The reaction was cooled to 0 °C and then iodomethane (115 ⁇ L, 1.85 mmol) was added.
  • the reaction turned a lighter orange colour upon addition.
  • the reaction was warmed to room temperature and left to stir overnight.
  • the reaction was quenched with a 1M aqueous solution of HCI and then extracted with EtOAc (x3).
  • the combined organic layers were washed with water then brine, dried over MgSO 4 , filtered and then concentrated under reduced pressure.
  • the crude material was adsorbed onto Celite® and purified by normal-phase silica gel chromatography using a 0% to 100% EtOAc in hexanes gradient.
  • the desired fractions were collected and concentrated under reduced pressure to afford 2-methyl-2-[1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]propanenitrile (134 mg, 69% yield) as an orange-brown oil.
  • Step 5 To a 20 mL scintillation vial was added 2-methyl-2-[1-(2- trimethylsilylethoxymethyl)benzimidazol-4-yl]propanenitrile (134 mg, 0.426 mmol) from the previous step followed by DCM (2 mL) and TFA (2 mL). The reaction was left to stir at room temperature overnight. The solvent was removed under reduced pressure. The material was dissolved in MeOH and then Amberlite IRA-67 resin was added. The resin was filtered out through a cotton plug.
  • Step 6 Preparation of the methylsulfone and methylsulfoxide mixture from an appropriate intermediate A-11 was performed as described above in Method B for Example 28 (step 1).
  • Step 7 Preparation of Examples 104, 112, 116-118 was performed by coupling the benzimidazole described in step 5 to the crude methylsulfone/methylsulfoxide mixture from A-11 (possessing an appropriate Ar group) using DI PEA in DMSO as described for Example 4 (step 7) in general method A.
  • Step 1 Preparation of 4-methoxy-1 H-imidazo[4,5-c]pyridine: In a pressure resistant tube, sodium hydride (104 mg, 2.60 mmol) was slowly added at 0 °C to 3.3 mL of MeOH. The resulting mixture was stirred for 5 min and commercial 4-chloro-1 H-imidazo[4,5-c]pyridine (0.10 g, 0.65 mmol) was added. The tube was sealed and the reaction mixture was heated at 120 °C and stirred for 12 hours. After cooling to room temperature, the resulting mixture was concentrated under reduced pressure. The crude product was purified by silica gel chromatography using a 0% to 10% MeOH in DCM gradient.
  • Step 1 Preparation of 3-ethoxy-2-nitro-aniline: A solution of sodium ethoxide (21 % in EtOH, 1.63 mL, 4.36 mmol) was added to a solution of 3-fluoro-2-nitro-aniline (0.23 g, 1 .45 mmol) in EtOH (8 mL). The resulting mixture was stirred at 80 °C for 10h. The reaction mixture was concentrated and water was added. The aqueous mixture was extracted with EtOAc. The organic layers were combined, washed with brine, dried with Na 2 SO 4 , filtered and concentrated.
  • Step 2 Preparation of 4-ethoxy-1 H-benzimidazole: Iron (0.37 g, 6.70 mmol) and ammonium chloride (0.36 g, 6.70 mmol) were added to a mixture of 3-ethoxy-2-nitro-aniline (0.24 g, 1.34 mmol) from the previous step in iPrOH (4.0 mL) and formic acid (1 .9 mL, 49.6 mmol). The resulting mixture was heated at 90 °C and stirred for 10h. The reaction mixture was cooled down to rt and filtered through Celite®.
  • Step 1 Preparation of tert-butyl 3-iodoindole-1 -carboxylate: Indole (1.00 g, 8.54 mmol) was dissolved in 26 mL of DMF in a 200 mL flask. Iodine (2.38 g, 9.39 mmol) and potassium hydroxide (1.20 g, 21.3 mmol) were then added. The reaction was stirred at room for 5 hours. It was then added to 100 mL of a saturated aqueous solution of Na 2 SO 3 . The mixture was then extracted with EtOAc three times. The combined organic layers were washed with water then brine, dried over MgSO 4 , filtered and then concentrated under reduced pressure.
  • Step 2 Preparation of 1-(1 H-indol-3-yl)pyrrolidin-2-one and tert-butyl 3-(2-oxopyrrolidin-1- yl)indole-1 -carboxylate: A flask was loaded with tert-butyl 3-iodoindole-1 -carboxylate (100 mg, 0.29 mmol), 2-pyrrolidinone (74 mg, 0.87 mmol), cesium carbonate (0.285 g, 0.87 mmol, Cui (28 mg, 0.15 mmol) and N,N"-dimethylethylenenediamine (31 ⁇ L, 0.29 mmol) in dioxane (1.5 mL).
  • the resulting mixture was stirred at 105 °C for a total of 3 days.
  • the mixture was cooled to room temperature then quenched with 0.2 mL of glacial acetic acid.
  • the product was isolated by prep HPLC using a 55% to 85% methanol in water gradient (with 0.1 % formic acid modifier) over 15 minutes.
  • Step 2 Preparation of the substituted benzimidazoles from the 2-nitroanilines described in step 1 was performed using the “one-pot” procedure described in Method G for Example 113 (step 2).
  • Step 1 Preparation of 1 H-benzo[d]imidazole-4-carbonitrile: To a nitrogen-purged (x3) 10 mL microwave vial equipped with a magnetic stir bar, was added 4-bromo-1 H-benzo[d]imidazole (99 mg, 0.502 mmol), dicyanozinc (70.8 mg, 0.603 mmol), palladium tetrakistriphenylphosphine (116 mg, 0.100 mmol) in DMF (4 mL). The reaction was heated to 90 °C and allowed to stir for 16h.
  • reaction was cooled to room temperature, diluted with water and then extracted with EtOAc.
  • Step 1 The synthesis of 2-[1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]acetonitrile was described for the synthesis of Examples 104, 112, 116-118.
  • Sodium hydride (100 mg, 2.51 mmol) was added to anhydrous DMF (5 mL) in a 20 mL vial. The mixture was cooled to 0 °C.
  • 2-[1-(2- trimethylsilylethoxymethyl)benzimidazol-4-yl]acetonitrile (240 mg, 0.84 mmol) in 5 mL of DMF was added slowly. The reaction was warmed to room temperature then was cooled back down to 0 °C.
  • Step 2 1-[1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]cyclopropanecarbonitrile (118 mg, 0.376 mmol) was dissolved in a mixture of DCM (2 mL) and TFA (2 mL) in a 20 mL vial. The reaction was left to stir at room temperature overnight. The solvent was removed under reduced pressure. The material was dissolved in MeOH and then Amberlite IRA-67 resin was added. The resin was filtered out and then the filtrate was concentrated under reduced pressure to afford 1- (1 H-benzimidazol-4-yl)cyclopropanecarbonitrile; 2,2,2-trifluoroacetic acid (114 mg, 102% yield) as a light brown, sticky solid.
  • Step 1 To a 100 mL round-bottom flask equipped with a magnetic stir bar was added DMF (25 mL) followed by 1H-benzo[d]imidazole-7-carboxylic acid (1.018 g, 6.28 mmol), EDCI (2.407 g, 12.56 mmol) , HOBT-H 2 O (1.923 g, 12.56 mmol) and triethylamine (1.750 ml, 12.56 mmol). The reaction was cooled to 0 °C and allowed to stir for 2 h. Next, concentrated ammonium hydroxide (2 ml, 29.6 mmol) was added and the reaction was allowed to warm to room temperature and stirred for 24 hours.
  • DMF 25 mL
  • 1H-benzo[d]imidazole-7-carboxylic acid 1.018 g, 6.28 mmol
  • EDCI 2.407 g, 12.56 mmol
  • HOBT-H 2 O 1.923 g, 12.56
  • Step 1 Cesium carbonate (8.35 g, 25.6 mmol) was charged in a 50 mL flask and suspended in 6 mL of DMSO. Dimethyl malonate (2.7 mL, 30.7 mmol) was then added at room temperature. After 10 minutes of stirring the thick slurry, 3-fluoro-2-nitro-aniline (800 mg, 5.12 mmol) was added affording a bright orange to red mixture which was stirred at 70 °C for 1 hour. The mixture was allowed to cool to room temperature then diluted with EtOAc and washed with a saturated solution of ammonium chloride.
  • the bright red mixture was heated at 150 °C for 2.5 hours after which time the color had become a bit darker.
  • the mixture was allowed to cool to room temperature then diluted with water and EtOAc.
  • the biphasic mixture was filtered through a short pad of Celite® to remove insolubles creating emulsions and the layers were separated.
  • the aqueous layer was further extracted 3 more times with EtOAc.
  • the combined organic layers were then washed with water twice and once with brine then dried over MgSO 4 , filtered and concentrated.
  • the bright red residue was purified by flash chromatography on silica gel using a 15% to 50% EtOAc in hexanes gradient. The appropriate fractions were pooled, concentrated and dried under reduced pressure.
  • Step 3 Methyl 2-(3-amino-2-nitro-phenyl)acetate (545 mg, 2.59 mmol) was charged in a 100 mL flask containing a magnetic stir bar. 5% Palladium on charcoal (109 mg, 0.051 mmol) was then added and the mixture was suspended in 14 mL of methanol. Triethyl orthoformate (0.91 mL, 5.45 mmol) was added followed by 2 drops of acetic acid. The reaction flask was placed under reduced pressure then hydrogen was introduced. This operation was repeated twice more then the mixture was stirred vigorously at room temperature for 24 hours under a balloon atmosphere of hydrogen.
  • Step 4 A solution of methyl 2-(1 H-benzimidazol-4-yl)acetate (40 mg, 0.21 mmol) in 2 mL of anhydrous THF was cooled in an ice/water bath and allowed to stir for 5 minutes. A 1M THF solution of lithium aluminum hydride (0.25 mL, 0.25 mmol) was then added dropwise. Some gas evolution was noticed and the clear yellow solution became a milky beige suspension. The mixture was allowed to warm to room temperature and stirred overnight. 3-4 Drops of a saturated ammonium chloride solution were added to the mixture. After 5 minutes of stirring, 100 mg of sodium sulfate decahydrate was added.
  • Example 133 Step 1 : To a 100 mL round-bottom flask, was added 4-bromo-1 H-benzimidazole (1.00 g, 5.08 mmol), 2-(trimethylsilyl)ethoxymethyl chloride (1.1 mL, 6.09 mmol ) and 60% NaH (244 mg, 6.09 mmol) in anhydrous DMF (10 mL). The reaction was allowed to stir at room temperature for 1h. Upon completion, the reaction was quenched with sat. NH 4 CI and then extracted EtOAc (x3). The organics were collected washed with brine, separated then dried by MgSO 4 , filtered and then concentrated under reduced pressure.
  • Step 2 To a flame-dried 5 mL microwave vial, was added 2-[(4-bromobenzimidazol-1- yl)methoxy]ethyl-trimethyl-silane (500 mg, 1.53 mmol) and bis(pinacolato)diboron (776 mg, 3.06 mmol) in DMF (0.30 mL) under a N 2 atmosphere. Next, 1 ,1'-bis(diphenylphosphino)ferrocene- palladium(l l)dichloride dichloromethane complex (279 mg, 0.382 mmol) and potassium acetate (450 mg, 4.58 mmol) were added to the solution. The reaction was sparged with a balloon of argon for a few minutes, sealed and then heated to 100 °C and left to stir for 16h. The reaction was cooled to room temperature. The crude reaction mixture was used as such in the next step.
  • Step 3 To the crude reaction mixture from step 2, were added 4-amino-3-bromopyridine (34 mg, 0.20 mmol), tetrakis(triphenylphosphine)palladium(0) (35 mg, 0.0305 mmol) and sodium carbonate (97 mg, 0.914 mmol). The reaction was sparged with a balloon of argon for ⁇ 5-10min, sealed and then heated to 100 °C for 24 hours. Upon completion, the reaction was cooled to room temperature. The reaction was diluted with brine and extracted with EtOAc (x3). The organic layers were collected, dried over MgSO 4 , filtered and then concentrated under reduced pressure.
  • 4-amino-3-bromopyridine 34 mg, 0.20 mmol
  • tetrakis(triphenylphosphine)palladium(0) 35 mg, 0.0305 mmol
  • sodium carbonate 97 mg, 0.914 mmol
  • Step 4 To a 20 mL scintillation vial, was added 3-[1-(2-trimethylsilylethoxymethyl)-benzimidazol- 4-yl]pyridin-4-amine (72 mg, 0.210 mmol) in DCM (2 mL) and TFA (2 mL). The reaction was left to stir at room temperature for 16h. Upon completion, the solvent was removed under reduced pressure and then dried under vacuum to afford 3-(1 H-benzimidazol-4-yl)pyridin-4-amine TFA salt (68 mg, 100 % yield) as a brown oil. MS m/z 211.2 (MIT).
  • Preparation of Example 134 Preparation of Example 134:
  • Step 1 To a 20 mL scintillation vial, was added 4-bromo-1 H-benzimidazole (500 mg, 2.54 mmol), 60% NaH (122 mg, 3.05 mmol) and 4-methoxybenzyl chloride (413 ⁇ L, 3.05 mmol) in DMF (5 mL). The reaction was allowed to stir at room temperature for 2h. The reaction was quenched with sat. NH 4 CI and then extracted with EtOAc (x3). The organics were collected, dried over MgSO 4 , filtered and then concentrated under reduced pressure.
  • Step 2 To a flame-dried 5 mL microwave vial was added 4-bromo-1-[(4- methoxyphenyl)methyl]benzimidazole from step 1 (200 mg, 0.63 mmol) and bis(pinacolato)diboron (320 mg, 1.26 mmol) in DMF (0.30 mL) under a N2 atmosphere. Next, 1,1'- bis(diphenylphosphino)ferrocene-palladium(ii)dichloride dichloromethane complex (231 mg, 0.315 mmol) and then potassium acetate (186 mg, 1.89 mmol) was added to the solution.
  • the reaction was sparged with a balloon of argon for a few minutes, sealed and then heated to 100 °C and left to stir for 24h.
  • the reaction was cooled to room temperature.
  • the reaction was extracted diluted with brine and extracted with EtOAc (x3).
  • the organics were collected, dried by MgSO 4 , filtered through a plug of Celite® and then concentrated under reduced pressure.
  • the crude material was loaded onto Celite® and then purified by silica gel chromatography with EtOAc in hexanes.
  • Step 3 To a flame-dried 5mL microwave vial, was added 1-[(4-methoxyphenyl)methyl]-4-(4, 4,5,5- tetramethyl-1 ,3,2-dioxaborolan-2-yl)benzimidazole (56 mg, 0.154 mmol) and 2-amino-3- bromopyridine (35 mg, 0.200 mmol) under a N2 atmosphere. Next, sodium carbonate (49 mg, 0.461 mmol) and then tetrakis(triphenylphosphine)palladium(0) (18 mg, 0.0154 mmol) were added to the solution.
  • Step 4 To a 20mL scintillation vial, was added 3-[1-[(4-methoxyphenyl)methyl]-benzimidazol-4- yl]pyridin-2-amine (15 mg, 0.0445 mmol) in TFA (2 mL). The reaction was heated to 80 °C and stirred for 96 hours. The solvent was removed under reduced pressure and then the product was co-evaporated with Toluene (x3) to afford 3-(1 H-benzimidazol-4-yl)pyridin-2-amine TFA salt (14 mg, 100%). MS m/z 211.2 (MH + ).
  • Step 1 Methyl 2-(1H-benzimidazol-4-yl)acetate prepared as described in step 3 of Example 126 (303 mg, 1.59 mmol) was dissolved in 10 mL of DMF then potassium carbonate (661 mg, 4.78 mmol) was added followed by SEM-CI (0.42 mL, 2.39 mmol) dropwise over 10 minutes at room temperature. The mixture was allowed to stir at the same temperature for 16 hours. The mixture was poured into a saturated solution of ammonium chloride then extracted 3x with EtOAc. The combined organic layers were washed with water then brine then dried over MgSO 4 filtered and concentrated.
  • Step 2 A mixture of methyl 2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1 H-benzo[d]imidazol-4- yl)acetate and methyl 2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1 H-benzo[d]imidazol-7-yl)acetate (265 mg, 0.83 mmol) was charged in a 25 mL flask.
  • Step 3 A solution of the methyl esters from step 2 (275 mg, 0.79 mmol) in 4 mL of anhydrous THF was cooled in an ice/water bath and allowed to stir for 5 minutes. A 1 M THF solution of lithium aluminum hydride (0.95 mL, 0.95 mmol) was then added dropwise. Some gas evolution was noticed. The mixture was allowed to warm to room temperature and stirred for 23 hours. Sodium sulfate decahydrate (0.7g) was added to the mixture which was allowed to stir for another hour. The mixture was then diluted with EtOAc and filtered through a pad of Celite®, rinsing with EtOAc.
  • the second product to elute is [1-(1 H-benzimidazol-4-yl)cyclopropyl]methanol (47 mg, 31 % yield). MS m/z 186.9 (M-H’).
  • Step 2 The methylsulfone/methylsulfoxide mixture from step 1 (82 mg, 0.16 mmol) was suspended in 1.3 mL of 95% EtOH and 0.1 mL of water. A 1M solution of hydrazine (0.77 mL, 0.77 mmol) in THF was then added. The resulting mixture was stirred at 45 °C for 2 hours. The mixture (which had become a pale beige suspension) was allowed to cool to room temperature then diluted with 10 mL of water. The solids from the resulting suspension were collected on a hardened paper filter in a Buchner funnel (very slow filtration). The solids were washed with a bit of water then dried under reduced pressure.
  • Step 3 Isopentyl nitrite (0.038 mL, 0.28 mmol) was charged in a 25 mL flask then diluted with 4 mL of acetonitrile. Copper dibromide (50.7 mg, 0.23 mmol) was then added and the dark green suspension was degassed by bubbling argon through it for 5 minutes then for another minute with sonication.
  • N-[2,4-difluoro-3-[(2-hydrazinothiazolo[5,4-d]pyrimidin-7-yl)amino]phenyl]-3-fluoro-2- methyl-benzenesulfonamide (91 mg, 0.19 mmol) (from step 2) was then added in 4 small portions (some gas evolution was noticed) at room temperature. Once the addition was complete, the mixture was allowed to stir for 20 minutes. It was then diluted with EtOAc and washed with water containing 3-4 mL of a saturated aqueous EDTA solution. After separation of the layers, the wash was repeated twice more (until no more blue color was present in the aqueous layer). The organic layer was then washed with water and then brine.
  • Step 4 Preparation of Example 154: N-[3-[(2-bromothiazolo[5,4-d]pyrimidin-7-yl)amino]-2,4- difluoro-phenyl]-3-fluoro-2-methyl-benzenesulfonamide (36 mg, 0.07 mmol) from step 3 was charged in 10 mL flask along with tributyl(thiazol-5-yl)stannane (38 mg, 0.10 mmol) and a magnetic stir bar. DMF (2 mL) was added and argon was bubbled through the mixture for 3-4 minutes. Tetrakis(triphenylphosphine)palladium(0) (7.8 mg, 0.007 mmol) was added.
  • Step 1 Preparation of Example 156: The precursor tert-butyl 4-(1-(7-((3-((2,3- dichlorophenyl)sulfonamido)-2,6-difluorophenyl)amino)thiazolo[5,4-d]pyrimidin-2-yl)-1 H- benzo[d]imidazol-4-yl)piperazine-1 -carboxylate was prepared as described in general method G for Example 113 using Boc-protected piperazine for the first step. This precursor (63 mg, 0.08 mmol) was dissolved in 0.8 mL of DCM and TFA (0.061 mL, 0.80 mmol) was added at room temperature.
  • Step 1 the starting 4-thiomorpholinebenzimidazole fragment was prepared following method G. A solution of oxone® (1.42 g, 2.32 mmol) in water (10 mL) was added to a solution of 4-(1 H- benzimidazol-4-yl)thiomorpholine (0.127 g, 0.579 mmol) in methanol (20 mL). The resulting mixture was stirred at rt for 17 h.
  • Example 159 Step 1 : Carbomethoxylation of 2,4,5-trifluoroaniline was performed as described in patent W02020/261156.
  • Step 2 To a solution of methyl 3-amino-2,5,6-trifluorobenzoate (2.72 g, 13.26 mmol) in DCE/pyridine (1 :1 , 16 mL) was added 2,3-dichlorobenzenesulfonyl chloride (3.91 g, 15.91 mmol) portion wise at rt. The reaction was heated to 70 °C for 16h. The reaction was monitored by LCMS. Upon completion, it was quenched with HCI 1 M. The aqueous layer was extracted with EtOAc (15 mL) three times. The combined organic layers were washed with brine, dried over MgSO 4 , filtered and evaporated to dryness.
  • Step 4 To a solution of 3-((2,3-dichlorophenyl)sulfonamido)-2,5,6-trifluorobenzoic acid from step 3 (4.50 g, 11.24 mmol) in acetonitrile (30 mL) was added triethylamine (1.71 mL, 12.37 mmol) and diphenyl phosphoryl azide (2.91 mL, 13.50 mmol) at rt. The reaction was heated to 80 °C overnight. The reaction was cooled down to rt and water (30 mL) was added. The aqueous layer was extracted with EtOAc (30 mL) three times.
  • Step 5 To a solution of 2,3-dichloro-N-(2,4,5-trifluoro-3-isocyanatophenyl)benzenesulfonamide from step 4 (1.35 g, 3.39 mmol) in THF (17 mL) was added aqueous LiOH 4M (17 mL) . The pressure vessel was sealed and heated to 100 °C for 1 h in an oil bath. When the reaction was completed, it was quenched with a saturated NH 4 CI solution. EtOAc was added and the layers were allowed to separate. The aqueous layer was extracted with EtOAc (20 mL) two more times.
  • Step 6 N-(3-amino-2,4,5-trifluoro-phenyl)-2,3-dichloro-benzenesulfonamide (300 mg, 0.81 mmol) from step 5 and 7-chloro-2-methylsulfanyl-thiazolo[5,4-d]pyrimidine (A-10, 194 mg, 0.89 mmol) were added to glacial AcOH (3.2 mL). The reaction was heated at 65 °C overnight and at 85 °C for 2 hours. Another portion of 7-chloro-2-methylsulfanyl-thiazolo[5,4-d]pyrimidine (A-10, 100 mg, 0.46 mmol) was added and the mixture was stirred at 100 °C for 24 hours.
  • Step 7 To Crude 2,3-dichloro-N-[2,4,5-trifluoro-3-[(2-methylsulfanylthiazolo[5,4-d]pyrimidin-7- yl)amino]phenyl]benzenesulfonamide (238 mg, 0.43 mmol) from step 6 in DCM (5 mL) was added 3-chloroperoxybenzoic acid (149 mg, 0.862 mmol). The resulting reaction mixture was allowed to stir at room temperature overnight then diluted with EtOAc and washed with a saturated aqueous solution of NaHCO 3 followed by brine. The organic layer was dried over MgSO 4 , filtered and concentrated. The resulting crude product (mostly methylsulfone, MS m/z 584.2 (MH + )) was carried through to the next step without further purification (66 mg, 26% yield).
  • Step 8 Crude 2,3-dichloro-N-[2,4,5-trifluoro-3-[(2-methylsulfonylthiazolo[5,4-d]pyrimidin-7- yl)amino]phenyl]benzenesulfonamide (66 mg, 0.11 mmol) was dissolved in 1.1 mL of DMSO. Benzimidazole (13 mg, 0.11 mmol) and cesium carbonate (74 mg, 0.23 mmol) were added and the reaction mixture was stirred at 100 °C overnight. After cooling, the reaction mixture was filtered through a short pad of silica gel and Celite®.
  • the filtrate was purified by reverse-phase chromatography using a methanol in water gradient with 0.1% formic acid as modifier. The appropriate fractions were pooled and concentrated. The residue was lyophilized from an acetonitrile/water mixture. Affords N-[3-[[2-(benzimidazol-1-yl)thiazolo[5,4-d]pyrimidin-7- yl]amino]-2,4,5-trifluoro-phenyl]-2,3-dichloro-benzenesulfonamide (Example 159, 12 mg, 17 % yield) as a white fluffy powder.
  • BRAF and CRAF In vitro enzymatic reactions were used for evaluating compounds intrinsic activity against BRAF, CRAF and ARAF.
  • BRAF and CRAF 0.375 nM of purified GST-tagged kinases (cat. No. B4062-10UG and cat. No. R1656-10UG respectively from Millipore Sigma) were incubated with 75nM of kinase-dead MEK1 substrate (cat. No. 40075; BPS Bioscience) in the presence of 10 ⁇ M of Ultrapure ATP (cat. No.
  • test substance solution 5 ⁇ L/well of test substance solution are placed in a 384-well proxyplate (Perkin Elmer) and mixed with 2x concentrated kinase reactions.
  • the dilution series is selected so that nine concentrations cover a range from 100 nM to 0.01 nM. If necessary (if the compound exhibits low intrinsic potency) the initial concentration of 100 nM is changed to 1 ⁇ M, or 0.5 ⁇ M and further dilutions are carried out accordingly.
  • the final concentration of DMSO in the assay is set at 0.05%.
  • BRAF and CRAF kinase reactions were carried out for a total of 2 hours at 30 °C and then stopped by 1/2 dilution in ADP-Glo Reagent (cat. No. V9102; Promega; part V912C). Reactions were then incubated for 1 h at room temperature before addition of one volume of Kinase Detection Reagent (cat. No. V9102; Promega; part V917A). Plates were then equilibrated at room temperature for 30 minutes before detection of luminescence on a Synergy Neo2 plate reader (Biotek). The effect of each compound dilution on BRAF and CRAF kinase activity was expressed as %inhibition and calculated as follows.
  • ARAF kinase reactions were carried out for a total of 2 hours at 30 °C and then stopped by addition of EDTA at a final concentration of 40mM. Reactions were then detected using the AlphaLISA® SureFire® UltraTM p-MEK 1/2 (Ser218/222) (PerkinElmer) kit. Reactions were performed with 5 ⁇ L of kinase reaction according to the manufacturer’s specifications in 384-well proxyplates (Perkin Elmer) followed by overnight incubation of the AlphaLISA® reactions at room temperature in a humidified chamber. After completion of the detection reactions, the signals were recorded on a Synergy Neo2 plate reader (Biotek) equipped with AlphaLISA® filters.
  • IC 50 values were obtained by plotting the kinase inhibition values and fitting the dose-activity curves using a log(agonist) versus response - variable slope (four parameters) function using either GraphPadPrism (V7.0) or Dotmatics Screening Ultra platform.
  • Standards included in the ARAF kinase assay were Belvarafenib (MedChem Express cat. No. HY-109080; CAS No. 1446113-23-0), LXH254 (MedChem Express cat. No. HY-112089; CAS No. 1800398-38-2) and BGB283 (cat. No. HY-18957; CAS No. 1446090-79-4).
  • BRAF, CRAF and ARAF ATP-competitive kinase inhibitors as demonstrated by direct inhibition of enzymatic activity in vitro.
  • BRAF and CRAF inhibition potencies of compounds are listed in Tables 3 and 4 while ARAF kinase inhibition potencies of representative analogs are listed in Table A.
  • All cancer cell lines (A375, A101 D, A2058, RKO, HT29 SK-MEL 30, IPC298, HepG2, HCT-116, Lovo, SW620, SW480, NCI-H358, NCI-H2 122, Calu-6, NCIH2087, NCIH1755, NCIH1666 and Mewo) were obtained from ATCC and cultured in RPMI-1640 medium (Gibco) supplemented with 5% heat inactivated fetal bovine serum (FBS, Wisent) at 37 °C under 5% CO 2 . Cells were maintained in T175 flasks (Greiner).
  • Table B Tumor type and RAS-ERK pathway mutational status of cancer cell lines (CCLs) used for pERK and antiproliferative profiling of substances described in this application.
  • AlphaLISA® SureFire® UltraTM p-ERK 1/2 (Thr202/Tyr204) analysis was conducted on cells plated in 100 ⁇ L of complete RPMI-1640 growth medium in 96-well flat-bottomed transparent dishes (Costar) at a density indicated in Table C. Cells were maintained overnight at 37 °C under 5% CO 2 before treatment with compounds’ dilution series for one hour. The cell density in cells/cm 2 corresponds to cell number divided by the area of one well of a 96-well plate (0.143 cm 2 ).
  • test substance dilution prepared in complete RPMI-1640 growth media was added to the cells.
  • the dilution series is selected so that ten concentrations cover a range from 10 ⁇ M to 0.33 nM. If necessary, the initial concentration of 10 ⁇ M is increased to 100 ⁇ M or lowered to 1 ⁇ M (as in the case of A375 and H1666 cells, which are generally more sensitive to the compounds) and further dilution is carried out accordingly.
  • the final concentration of DMSO in the assay is set at 0.5%.
  • %inhibition 100*(1-((pERK signal compound )/ (pERK signal DMSO )))
  • the ability of each compound to inhibit pERK signal was expressed as an IC 50 value obtained by plotting the inhibition values for each data point of a dilution series and fitting the obtained curves using a log(agonist) versus response - variable slope (four parameters) function using GraphPadPrism (V7.0) or Dotmatics Screening Ultra platform.
  • %Y MIN of IC 50 curves was greater than -20%; where Y MIN corresponded to the data point having the lowest value in the IC 50 curve of the said compound.
  • Figure 1 provides a visualization of the IC 50 curves for a compound that induces paradoxical pathway activation (PLX4720, commercially available from Selleck Chemicals; CAS No. 918505-84-7) and representative compounds as described herein exhibiting the unexpected and distinct induction-free profile.
  • Figure 1 shows representative IC 50 inhibition dose response curves for compounds as described herein that do not induce paradoxical induction of pERK signaling (Y MIN >-20%) in RAS-mutant HCT116 cells (Examples 44 and 122) and a compound (PLX4720) that causes strong induction of the pathway in the same cell line (Y MIN ⁇ -600%).
  • the present compounds do not induce paradoxical activation of the pathway according to the criteria described above.
  • Example compounds 1 to 159 show pERK inhibition activity in the colon G13D Ras-mutated HCT- 116 cell line as shown in Tables 3 and 4. In addition, some Examples were also shown to display paradoxical induction-free inhibition of pERK signaling in the SW480 colon cell line harboring the G12D allele of KRAS (Tables 3 and 4). Furthermore, some Examples from Tables 3 and 4 were also tested for inhibition of pERK in A375 cells that comprise the BRAF V600E driver mutation and were found to be active as well (Table D-1 and D-2). All compound Examples 1-159 displayed pERK IC 50 values in the HCT 116 cell line that were ⁇ 30 ⁇ M.
  • Representative compounds as herein defined were also tested on additional tumor cells for their pERK inhibition activity and showed good to very good pERK inhibition activity in cancer cell lines bearing various NRAS, KRAS and NF1 alleles and representative of a large diversity of tissue types (i.e., SK-MEL 30, IPC298, HepG2, HCT-116, Lovo, SW620, SW480, NCI-H358, NCI- H2122, Calu-6 and Mewo; see Table D-1 and D-2 and refer to Table B for genotypes).
  • tissue types i.e., SK-MEL 30, IPC298, HepG2, HCT-116, Lovo, SW620, SW480, NCI-H358, NCI- H2122, Calu-6 and Mewo; see Table D-1 and D-2 and refer to Table B for genotypes).
  • the pERK inhibition activity of compounds is stronger in cancer cell lines bearing various BRAF alleles (A375, A101 D, A2058, RKO, HT29, NCIH2087, NCI H 1755 and NCI H 1666) (Table D-1 and D-2).
  • Table D (D-1 and D-2). Induction-free pERK IC 50 values and anti-proliferative EC 50 values for select compounds in a panel of RAS-mutant cancer cell lines (see Table B for genotypes) and BRAF V600E mutant A375.
  • the %Y m in values for pERK IC 50 curves were all above -20% and considered to display minimal or no induction and thus compounds do not cause detectable paradoxical activation of the pathway in this panel of cancer cell lines.
  • the comparative results for the molecule Belvarafenib (obtained from MedChem Express cat. No. HY-109080; CAS No. 1446113-23-0) causes mild to strong induction of the pathway in the same cell lines (Y MIN ⁇ - 30% in 10 of the 13 RAS-mutant cell lines tested).
  • CellTiter-Glo® viability analysis was conducted on cells plated in 100 ⁇ L of complete RPMI-1640 growth medium in 96-well flat-bottomed white opaque plates (Greiner or Croning) at a density indicated in Table E (for each CCL, number of cells plated per well of a 96-well plate to perform the CellTiter-Glo® cell viability assay).
  • the cell density in cells/cm 2 would correspond to cell number divided by the area of one well of a 96-well plate (0.32 cm 2 ). Cells were maintained overnight at 37 °C under 5% CO 2 before treatment with compounds’ dilution series for 3 days.
  • Table E Number of cells plated per well of a 96-well plate to perform the CellTiter-Glo® cell viability assay
  • test substance dilution prepared in complete RPMI-1640 growth media was added to the cells plated initially in 100 ⁇ L of growth media.
  • the dilution series is selected so that ten concentrations cover a range from 10 ⁇ M to 0.33 nM. If necessary (as in the case of A375 cells, which were more sensitive to the compounds), the initial concentration of 10 ⁇ M is lowered to 1 ⁇ M and further dilution is carried out accordingly.
  • the final concentration of DMSO in the assay is set to 0.5%.
  • each compound to inhibit proliferation was expressed as a EC 50 value obtained by plotting the effect values for each data point of a dilution series and fitting the obtained curves using a log(agonist) versus response - variable slope (four parameters) function using GraphPadPrism (V7.0) or Dotmatics Screening Ultra platform.
  • the active substances show antiproliferative activity in various NRAS-, KRAS- and NF1-mutant cancer cell lines that are representative of a large diversity of tissue types (i.e. , SK-MEL 30, IPC298, HepG2, HCT-116, Lovo, SW620, SW480, NCI-H358, NCI- H2122, Calu-6 and Mewo; Tables D-1 and D-2 and refer to Table B for genotypes).
  • Antiproliferative activity is often even stronger in cell lines carrying BRAF driver mutations (A375, A101 D, A2058, RKO, HT29, NCIH2087, NCIH1755 and NCIH1666) (Tables D-1 and D-2).
  • BRAF driver mutations A375, A101 D, A2058, RKO, HT29, NCIH2087, NCIH1755 and NCIH1666) (Tables D-1 and D-2).
  • ⁇ denotes an IC 50 >5O nM
  • ⁇ denotes a 10-50 nM IC 50 range
  • ⁇ denotes an IC 50 ⁇ 10 nM.
  • Example 159 Characterization of Example 159 from Table 4 Numerous modifications could be made to any of the embodiments described above without departing from the scope of the present invention. Any references, patents or scientific literature documents referred to in the present document are incorporated herein by reference in their entirety for all purposes.

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Abstract

L'invention concerne des composés, des compositions et leur utilisation dans le traitement d'une maladie ou d'un état prolifératif tel qu'une maladie ou un trouble prolifératif associé à une mutation du gène RAF et/ou une mutation du gène RAS. Les composés décrits sont de formule I ou un sel et/ou solvate pharmaceutiquement acceptable de ceux-ci, où R1, R2, X1, X2, X3 et X4 sont tels que définis dans la description : (Formule i)
PCT/CA2023/051356 2022-10-13 2023-10-12 Composés thiazolo[5,4-d]pyrimidine, compositions les comprenant et leurs utilisations WO2024077391A1 (fr)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010026262A1 (fr) * 2008-09-08 2010-03-11 Boehringer Ingelheim International Gmbh Pyrido-[5,4-d]-pyrimidines en tant qu’inhibiteurs de prolifération cellulaire
WO2012101238A1 (fr) * 2011-01-27 2012-08-02 Boehringer Ingelheim International Gmbh Nouveaux pyrimido[5,4-d]pyrimidylamino phényl sulfonamides utilisés comme inhibiteurs de la sérine/thréonine kinase
US20150158854A1 (en) * 2013-12-11 2015-06-11 Development Center For Biotechnology Quinazoline compounds, method for preparing the same and use thereof

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010026262A1 (fr) * 2008-09-08 2010-03-11 Boehringer Ingelheim International Gmbh Pyrido-[5,4-d]-pyrimidines en tant qu’inhibiteurs de prolifération cellulaire
WO2012101238A1 (fr) * 2011-01-27 2012-08-02 Boehringer Ingelheim International Gmbh Nouveaux pyrimido[5,4-d]pyrimidylamino phényl sulfonamides utilisés comme inhibiteurs de la sérine/thréonine kinase
US20150158854A1 (en) * 2013-12-11 2015-06-11 Development Center For Biotechnology Quinazoline compounds, method for preparing the same and use thereof

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